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1
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Nataraj BH, Ramesh C, Mallappa RH. Probiotic and postbiotic interference exhibit anti-adhesion effects against clinical methicillin-resistant Staphylococcus aureus (MRSA) and impede MRSA-induced intestinal epithelial hyper-permeability in HT-29 cell line. Microb Pathog 2025; 199:107215. [PMID: 39647539 DOI: 10.1016/j.micpath.2024.107215] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/15/2024] [Revised: 12/06/2024] [Accepted: 12/06/2024] [Indexed: 12/10/2024]
Abstract
This study investigates the dynamics of MRSA de-colonization on HT-29 cell line using effective strategies like probiotics and postbiotics. Exploring novel alternatives to combat infections caused by antibiotic-resistant pathogens is an urgent need. Harnessing the antagonistic properties of live probiotics and their heat-killed preparations (postbiotics) to curb the growth of AMR pathogens represents a promising and essential area of contemporary research. This study was designed to evaluate the anti-adhesion properties of indigenous probiotics (Limosilactobacillus fermentum Lf1 and Lactiplantibacillus plantarum A5), as well as standard reference strains (Lacticaseibacillus rhamnosus GG and Lactobacillus acidophilus NCFM), and their heat-killed postbiotic preparations against clinical MRSA isolates (MRSA12/206 and 5/255) on the HT-29 cell line. ATR-FTIR-based functional group characterization of the postbiotic preparations revealed the heat-induced alterations in cell surface molecules and architecture. Both probiotic and postbiotic preparations were non-cytotoxic to HT-29 cells. The probiotic intervention, via protective, competitive, and displacement modes, significantly (p < 0.05) reduced the adhesion of MRSA isolates to HT-29 cells, with the protective and competitive modes showing greater efficacy. In contrast, heat-killed probiotics demonstrated notable anti-MRSA adhesion effects across all three modes (protective, competitive, and displacement). In comparison, heat-killed cells exhibited a superior anti-adhesion capability compared to live cells, likely due to the enhanced accessibility of microbe-associated molecular patterns and adhesion sites following heat treatment. Furthermore, co-treatment of MRSA with probiotic strains substantially (p < 0.05) reduced FITC-dextran transflux across the HT-29 cell monolayer. In conclusion, this study highlights the superior anti-adhesion efficacy of heat-killed postbiotics over live probiotic cells against MRSA isolates. It underscores the further need for pre-clinical and in-vivo investigations to validate the anti-MRSA colonization and gut barrier prophylactic or therapeutic potential of the investigated probiotics and postbiotics. Thus, the present study documents and supports the alternative to antibiotics potential of probiotics and postbiotics.
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Affiliation(s)
- Basavaprabhu Haranahalli Nataraj
- Molecular Biology Unit, Dairy Microbiology Division, ICAR-National Dairy Research Institute, Karnal, 132001, Haryana, India; Dairy Bacteriology Section, Southern Regional Station, ICAR-National Dairy Research Institute, Adugodi, 560030, Bengaluru, Karnataka, India.
| | - Chette Ramesh
- Molecular Biology Unit, Dairy Microbiology Division, ICAR-National Dairy Research Institute, Karnal, 132001, Haryana, India.
| | - Rashmi Hogarehalli Mallappa
- Molecular Biology Unit, Dairy Microbiology Division, ICAR-National Dairy Research Institute, Karnal, 132001, Haryana, India; Dairy Bacteriology Section, Southern Regional Station, ICAR-National Dairy Research Institute, Adugodi, 560030, Bengaluru, Karnataka, India.
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2
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Chaudhry G, Zeenia, Safdar N, Begum S, Akim AM, Sung YY, Muhammad TST. Cytotoxicity assays for cancer drug screening: methodological insights and considerations for reliable assessment in drug discovery. BRAZ J BIOL 2024; 84:e284409. [PMID: 39699393 DOI: 10.1590/1519-6984.284409] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/12/2024] [Accepted: 09/19/2024] [Indexed: 12/20/2024] Open
Abstract
The importance of cytotoxicity assays in in vitro drug discovery investigations has led to their rising profile. Drugs and other substances can disrupt cell membranes, limit protein synthesis, and bind irreversibly to receptors, all of which lead to cell death in cancer cells. To precisely measure the cell death resulting from these damages, one must choose a cytotoxicity test that meets specific criteria. A systematic search strategy was used to gather grey literature from 2001 to 2024, utilizing databases such as PubMed and Google Scholar. Specific keywords related to colorimetric, fluorometric, and dye exclusion assays, as well as "cytotoxicity," were employed. Here, we only focus on screening drug cytotoxicity for cancer cells. This review discusses various cytotoxicity assays, such as "dye exclusion assays," "colorimetric assays," and "fluorometric assays." It is crucial to prioritize safety, speed, reliability, efficiency, and cost-effectiveness, while also ensuring minimal interference with the test compound. Commonly used in toxicology and pharmacology, cytotoxicity assays are based on several biological processes. Selecting the correct assay method requires considerations such as assay specificity and sensitivity, detection mechanism, test drug properties, and laboratory availability. This review aims to assist researchers in performing reliable cytotoxicity assessments by providing insights into assay choices.
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Affiliation(s)
- G Chaudhry
- Universiti Malaysia Terengganu, Institute of Climate Adaptation and Marine Biotechnology, Kuala Terengganu, Malaysia
| | - Zeenia
- Universiti Malaysia Terengganu, Institute of Climate Adaptation and Marine Biotechnology, Kuala Terengganu, Malaysia
| | - N Safdar
- Fatima Jinnah Women University, Department of Biotechnology, Rawalpindi, Pakistan
| | - S Begum
- Fatima Jinnah Women University, Environmental Sciences Department, Rawalpindi, Pakistan
| | - A M Akim
- Universiti Putra Malaysia, Department of Biomedical Sciences, Seri Kembangan, Selangor, Malaysia
| | - Y Y Sung
- Universiti Malaysia Terengganu, Institute of Climate Adaptation and Marine Biotechnology, Kuala Terengganu, Malaysia
| | - T S T Muhammad
- Universiti Malaysia Terengganu, Institute of Climate Adaptation and Marine Biotechnology, Kuala Terengganu, Malaysia
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3
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Liu Y, Wai AP, Zolzaya T, Iida Y, Okada S, Iizasa H, Yoshiyama H. Exploring the anti-EBV potential of suberoylanilide hydroxamic acid: Induction of apoptosis in infected cells through suppressing BART gene expression and inducing lytic infection. Virology 2024; 597:110161. [PMID: 38981317 DOI: 10.1016/j.virol.2024.110161] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/29/2024] [Revised: 06/09/2024] [Accepted: 06/24/2024] [Indexed: 07/11/2024]
Abstract
Epstein-Barr virus (EBV) is linked to lymphoma and epithelioma but lacks drugs specifically targeting EBV-positive tumors. BamHI A Rightward Transcript (BART) miRNAs are expressed in all EBV-positive tumors, suppressing both lytic infection and host cell apoptosis. We identified suberoylanilide hydroxamic acid (SAHA), an inhibitor of histone deacetylase enzymes, as an agent that suppresses BART promoter activity and transcription of BART miRNAs. SAHA treatment demonstrated a more pronounced inhibition of cell proliferation in EBV-positive cells compared to EBV-negative cells, affecting both p53 wild-type and mutant gastric epithelial cells. SAHA treatment enhanced lytic infection in wild-type EBV-infected cells, while also enhancing cell death in BZLF1-deficient EBV-infected cells. It reduced BART gene expression by 85% and increased the expression of proapoptotic factors targeted by BART miRNAs. These findings suggest that SAHA not only induces lytic infection but also leads to cell death by suppressing BART miRNA transcription and promoting the apoptotic program.
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Affiliation(s)
- Yuxin Liu
- Department of Microbiology, Faculty of Medicine, Shimane University, 89-1 Enya, Izumo, Shimane, 693-8501, Japan.
| | - Aung Phyo Wai
- Department of Microbiology, Faculty of Medicine, Shimane University, 89-1 Enya, Izumo, Shimane, 693-8501, Japan.
| | - Tumurgan Zolzaya
- Department of Microbiology, Faculty of Medicine, Shimane University, 89-1 Enya, Izumo, Shimane, 693-8501, Japan.
| | - Yuichi Iida
- Department of Immunology, Faculty of Medicine, Shimane University, 89-1 Enya, Izumo, Shimane, 693-8501, Japan.
| | - Shunpei Okada
- Department of Microbiology, Faculty of Medicine, Shimane University, 89-1 Enya, Izumo, Shimane, 693-8501, Japan.
| | - Hisashi Iizasa
- Department of Microbiology, Faculty of Medicine, Shimane University, 89-1 Enya, Izumo, Shimane, 693-8501, Japan.
| | - Hironori Yoshiyama
- Department of Microbiology, Faculty of Medicine, Shimane University, 89-1 Enya, Izumo, Shimane, 693-8501, Japan.
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4
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Camargo L, Gering I, Mastalipour M, Kraemer-Schulien V, Bujnicki T, Willbold D, Coronado MA, Eberle RJ. A Snake Venom Peptide and Its Derivatives Prevent Aβ 42 Aggregation and Eliminate Toxic Aβ 42 Aggregates In Vitro. ACS Chem Neurosci 2024; 15:2600-2611. [PMID: 38957957 PMCID: PMC11258689 DOI: 10.1021/acschemneuro.4c00089] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/07/2024] [Revised: 05/28/2024] [Accepted: 06/21/2024] [Indexed: 07/04/2024] Open
Abstract
Over a century has passed since Alois Alzheimer first described Alzheimer's disease (AD), and since then, researchers have made significant strides in understanding its pathology. One key feature of AD is the presence of amyloid-β (Aβ) peptides, which form amyloid plaques, and therefore, it is a primary target for treatment studies. Naturally occurring peptides have garnered attention for their potential pharmacological benefits, particularly in the central nervous system. In this study, nine peptide derivatives of Crotamine, a polypeptide from Crotalus durissus terrificus Rattlesnake venom, as well as one d-enantiomer, were evaluated for their ability to modulate Aβ42 aggregation through various assays such as ThT, QIAD, SPR, and sFIDA. All tested peptides were able to decrease Aβ42 aggregation and eliminate Aβ42 aggregates. Additionally, all of the peptides showed an affinity for Aβ42. This study is the first to describe the potential of crotamine derivative peptides against Aβ42 aggregation and to identify a promising d-peptide that could be used as an effective pharmacological tool against AD in the future.
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Affiliation(s)
- Luana
Cristina Camargo
- Institute
of Biological Information Processing (IBI-7: Structural Biochemistry), Forschungszentrum Jülich, Jülich 52428, Germany
- Faculty
of Mathematics and Natural Sciences, Institute of Physical Biology, Heinrich Heine University Düsseldorf, Düsseldorf 40225, Germany
| | - Ian Gering
- Institute
of Biological Information Processing (IBI-7: Structural Biochemistry), Forschungszentrum Jülich, Jülich 52428, Germany
| | - Mohammadamin Mastalipour
- Faculty
of Mathematics and Natural Sciences, Institute of Physical Biology, Heinrich Heine University Düsseldorf, Düsseldorf 40225, Germany
| | - Victoria Kraemer-Schulien
- Institute
of Biological Information Processing (IBI-7: Structural Biochemistry), Forschungszentrum Jülich, Jülich 52428, Germany
| | - Tuyen Bujnicki
- Institute
of Biological Information Processing (IBI-7: Structural Biochemistry), Forschungszentrum Jülich, Jülich 52428, Germany
| | - Dieter Willbold
- Institute
of Biological Information Processing (IBI-7: Structural Biochemistry), Forschungszentrum Jülich, Jülich 52428, Germany
- Faculty
of Mathematics and Natural Sciences, Institute of Physical Biology, Heinrich Heine University Düsseldorf, Düsseldorf 40225, Germany
| | - Mônika A. Coronado
- Institute
of Biological Information Processing (IBI-7: Structural Biochemistry), Forschungszentrum Jülich, Jülich 52428, Germany
- Faculty
of Mathematics and Natural Sciences, Institute of Physical Biology, Heinrich Heine University Düsseldorf, Düsseldorf 40225, Germany
| | - Raphael J. Eberle
- Institute
of Biological Information Processing (IBI-7: Structural Biochemistry), Forschungszentrum Jülich, Jülich 52428, Germany
- Faculty
of Mathematics and Natural Sciences, Institute of Physical Biology, Heinrich Heine University Düsseldorf, Düsseldorf 40225, Germany
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Welsh A, Matshitse R, Khan SF, Nyokong T, Prince S, Smith GS. Trinuclear ruthenium(II) polypyridyl complexes: Evaluation as photosensitizers for enhanced cervical cancer treatment. J Inorg Biochem 2024; 256:112545. [PMID: 38581803 DOI: 10.1016/j.jinorgbio.2024.112545] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/06/2023] [Revised: 03/23/2024] [Accepted: 03/24/2024] [Indexed: 04/08/2024]
Abstract
Trinuclear ruthenium(II) polypyridyl complexes anchored to benzimidazole-triazine / trisamine scaffolds were investigated as photosensitizers for photodynamic therapy. The trinuclear complexes were noted to produce a significant amount of singlet oxygen in both DMF and aqueous media, are photostable and show appreciable emission quantum yields (ɸem). In our experimental setting, despite the moderate phototoxic activity in the HeLa cervical cancer cell line, the phototoxic indices (PI) of the trinuclear complexes are superior relative to the PIs of a clinically approved photosensitizer, Photofrin®, and the pro-drug 5-aminolevulinic acid (PI: >7 relative to PI: >1 and PI: 4.4 for 5-aminolevulinic acid and Photofrin®, respectively). Furthermore, the ruthenium complexes were noted to show appreciable long-term cytotoxicity upon light irradiation in HeLa cells in a concentration-dependent manner. Consequently, this long-term activity of the ruthenium(II) polypyridyl complexes embodies their ability to reduce the probability of the recurrence of cervical cancer. Taken together, this presents a strong motivation for the development of polymetallic complexes as anticancer agents.
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Affiliation(s)
- Athi Welsh
- Department of Chemistry, University of Cape Town, Rondebosch 7700, ,South Africa
| | - Refilwe Matshitse
- Institute for Nanotechnology Innovation, Rhodes University, Makhanda 6140, South Africa
| | - Saif F Khan
- Division of Cell Biology, Department of Human Biology, University of Cape Town, Faculty of Health Science, Observatory, 7925, South Africa
| | - Tebello Nyokong
- Institute for Nanotechnology Innovation, Rhodes University, Makhanda 6140, South Africa
| | - Sharon Prince
- Division of Cell Biology, Department of Human Biology, University of Cape Town, Faculty of Health Science, Observatory, 7925, South Africa
| | - Gregory S Smith
- Department of Chemistry, University of Cape Town, Rondebosch 7700, ,South Africa.
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Khalef L, Lydia R, Filicia K, Moussa B. Cell viability and cytotoxicity assays: Biochemical elements and cellular compartments. Cell Biochem Funct 2024; 42:e4007. [PMID: 38593323 DOI: 10.1002/cbf.4007] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/10/2024] [Revised: 03/01/2024] [Accepted: 03/29/2024] [Indexed: 04/11/2024]
Abstract
Cell viability and cytotoxicity assays play a crucial role in drug screening and evaluating the cytotoxic effects of various chemicals. The quantification of cell viability and proliferation serves as the cornerstone for numerous in vitro assays that assess cellular responses to external factors. In the last decade, several studies have developed guidelines for defining and interpreting cell viability and cytotoxicity based on morphological, biochemical, and functional perspectives. As this domain continues to experience ongoing growth, revealing new mechanisms orchestrating diverse cell cytotoxicity pathways, we suggest a revised classification for multiple assays employed in evaluating cell viability and cell death. This classification is rooted in the cellular compartment and/or biochemical element involved, with a specific focus on mechanistic and essential aspects of the process. The assays are founded on diverse cell functions, encompassing metabolic activity, enzyme activity, cell membrane permeability and integrity, adenosine 5'-triphosphate content, cell adherence, reduction equivalents, dye inclusion or exclusion, constitutive protease activity, colony formation, DNA fragmentation and nuclear splitting. These assays present straightforward, reliable, sensitive, reproducible, cost-effective, and high-throughput approaches for appraising the effects of newly formulated chemotherapeutic biomolecules on the cell survival during the drug development process.
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Affiliation(s)
- Lefsih Khalef
- Département de Biochimie et Microbiologie, Laboratoire d'Ecologie, Biotechnologie et Santé, Université Mouloud Mammeri de Tizi ouzou, Tizi Ouzou, Algeria
| | - Radja Lydia
- Département de Biochimie et Microbiologie, Laboratoire d'Ecologie, Biotechnologie et Santé, Université Mouloud Mammeri de Tizi ouzou, Tizi Ouzou, Algeria
| | - Khettar Filicia
- Département de Biochimie et Microbiologie, Laboratoire d'Ecologie, Biotechnologie et Santé, Université Mouloud Mammeri de Tizi ouzou, Tizi Ouzou, Algeria
| | - Berkoud Moussa
- Département de Biochimie et Microbiologie, Laboratoire d'Ecologie, Biotechnologie et Santé, Université Mouloud Mammeri de Tizi ouzou, Tizi Ouzou, Algeria
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7
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Celis T, Bullens DMA, Hoet PHM, Ghosh M. Development and validation of a human bronchial epithelial spheroid model to study respiratory toxicity in vitro. Arch Toxicol 2024; 98:493-505. [PMID: 38148415 DOI: 10.1007/s00204-023-03619-9] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/29/2023] [Accepted: 10/04/2023] [Indexed: 12/28/2023]
Abstract
The use of laboratory animals in research has been extensively criticized. While most of the critique has been centered around the ethical aspect, also the economic and scientific aspects have been frequently mentioned as points of concern. As a result, the use of alternative methods has gradually become more enticing. The most used alternatives to laboratory animals are the 2D monolayer cell cultures. However, the limited translatability of these monolayer cell cultures to in vivo has led to the development of 3D cell cultures that are believed to better capture the in vivo physiology and pathology. Here we report on the development of a physiologically more relevant 3D cell model (spheroids) comprised of human bronchial epithelial (16HBE14o-) cells, for use in respiratory toxicity research. Culturing 16HBE14o-cells as hanging-drops led to the formation of stable spheroids which showed an increased expression of CLDN1 when compared to 2D monolayer cultured cells. In addition, cell-cycle analysis revealed an increased sub-G0 population and signs of G0/G1 arrest in spheroids. Afterwards, standard operating procedures (SOPs) were established, and existing protocols optimized, for compatibility with spheroids. Spheroids were successfully used to assess cytotoxicity, genotoxicity, apoptosis/necrosis, and oxidative stress after exposure to known cytotoxic or genotoxic compounds. The development of the bronchial epithelial spheroids and the establishment of SOPs can contribute to a more reliable toxicity assessment of chemicals and may aid in bridging the gap between in vivo and in vitro experiments.
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Affiliation(s)
- Thomas Celis
- Environment and Health Unit, Department of Public Health and Primary Care, KU Leuven, 3000, Leuven, Belgium
| | - Dominique M A Bullens
- Allergy and Clinical Immunology Research Group, Department of Microbiology, Immunology and Transplantation, KU Leuven, 3000, Leuven, Belgium
| | - Peter H M Hoet
- Environment and Health Unit, Department of Public Health and Primary Care, KU Leuven, 3000, Leuven, Belgium.
| | - Manosij Ghosh
- Environment and Health Unit, Department of Public Health and Primary Care, KU Leuven, 3000, Leuven, Belgium.
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8
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Wu CP, Murakami M, Li YC, Huang YH, Chang YT, Hung TH, Wu YS, Ambudkar SV. Imperatorin Restores Chemosensitivity of Multidrug-Resistant Cancer Cells by Antagonizing ABCG2-Mediated Drug Transport. Pharmaceuticals (Basel) 2023; 16:1595. [PMID: 38004460 PMCID: PMC10674403 DOI: 10.3390/ph16111595] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/02/2023] [Revised: 11/03/2023] [Accepted: 11/10/2023] [Indexed: 11/26/2023] Open
Abstract
The high expression of the ATP-binding cassette (ABC) drug transporter ABCG2 in cancer cells contributes to the emergence of multidrug resistance (MDR) in individuals afflicted with either solid tumors or blood cancers. MDR poses a major impediment in the realm of clinical cancer chemotherapy. Recently, substantial endeavors have been dedicated to identifying bioactive compounds isolated from nature capable of counteracting ABCG2-mediated MDR in cancer cells. Imperatorin, a natural coumarin derivative renowned for its diverse pharmacological properties, has not previously been explored for its impact on cancer drug resistance. This study investigates the chemosensitizing potential of imperatorin in ABCG2-overexpressing cancer cells. Experimental results reveal that at sub-toxic concentrations, imperatorin significantly antagonizes the activity of ABCG2 and reverses ABCG2-mediated MDR in a concentration-dependent manner. Furthermore, biochemical data and in silico analysis of imperatorin docking to the inward-open conformation of human ABCG2 indicate that imperatorin directly interacts with multiple residues situated within the transmembrane substrate-binding pocket of ABCG2. Taken together, these results furnish substantiation that imperatorin holds promise for further evaluation as a potent inhibitor of ABCG2, warranting exploration in combination drug therapy to enhance the effectiveness of therapeutic agents for patients afflicted with tumors that exhibit high levels of ABCG2.
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Affiliation(s)
- Chung-Pu Wu
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan; (Y.-C.L.); (Y.-H.H.)
- Department of Physiology and Pharmacology, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
- Department of Obstetrics and Gynecology, Taipei Chang Gung Memorial Hospital, Taipei 10507, Taiwan;
| | - Megumi Murakami
- Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA;
| | - Yen-Ching Li
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan; (Y.-C.L.); (Y.-H.H.)
| | - Yang-Hui Huang
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan; (Y.-C.L.); (Y.-H.H.)
| | - Yu-Tzu Chang
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan; (Y.-C.L.); (Y.-H.H.)
| | - Tai-Ho Hung
- Department of Obstetrics and Gynecology, Taipei Chang Gung Memorial Hospital, Taipei 10507, Taiwan;
- Department of Medicine, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
- Department of Obstetrics and Gynecology, Keelung Chang Gung Memorial Hospital, Keelung 20401, Taiwan
| | - Yu-Shan Wu
- Department of Chemistry, Tunghai University, Taichung 40704, Taiwan;
| | - Suresh V. Ambudkar
- Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA;
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9
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Corner TP, Teo RZR, Wu Y, Salah E, Nakashima Y, Fiorini G, Tumber A, Brasnett A, Holt-Martyn JP, Figg WD, Zhang X, Brewitz L, Schofield CJ. Structure-guided optimisation of N-hydroxythiazole-derived inhibitors of factor inhibiting hypoxia-inducible factor-α. Chem Sci 2023; 14:12098-12120. [PMID: 37969593 PMCID: PMC10631261 DOI: 10.1039/d3sc04253g] [Citation(s) in RCA: 8] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/14/2023] [Accepted: 10/12/2023] [Indexed: 11/17/2023] Open
Abstract
The human 2-oxoglutarate (2OG)- and Fe(ii)-dependent oxygenases factor inhibiting hypoxia-inducible factor-α (FIH) and HIF-α prolyl residue hydroxylases 1-3 (PHD1-3) regulate the response to hypoxia in humans via catalysing hydroxylation of the α-subunits of the hypoxia-inducible factors (HIFs). Small-molecule PHD inhibitors are used for anaemia treatment; by contrast, few selective inhibitors of FIH have been reported, despite their potential to regulate the hypoxic response, either alone or in combination with PHD inhibition. We report molecular, biophysical, and cellular evidence that the N-hydroxythiazole scaffold, reported to inhibit PHD2, is a useful broad spectrum 2OG oxygenase inhibitor scaffold, the inhibition potential of which can be tuned to achieve selective FIH inhibition. Structure-guided optimisation resulted in the discovery of N-hydroxythiazole derivatives that manifest substantially improved selectivity for FIH inhibition over PHD2 and other 2OG oxygenases, including Jumonji-C domain-containing protein 5 (∼25-fold), aspartate/asparagine-β-hydroxylase (>100-fold) and histone Nε-lysine demethylase 4A (>300-fold). The optimised N-hydroxythiazole-based FIH inhibitors modulate the expression of FIH-dependent HIF target genes and, consistent with reports that FIH regulates cellular metabolism, suppressed lipid accumulation in adipocytes. Crystallographic studies reveal that the N-hydroxythiazole derivatives compete with both 2OG and the substrate for binding to the FIH active site. Derivatisation of the N-hydroxythiazole scaffold has the potential to afford selective inhibitors for 2OG oxygenases other than FIH.
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Affiliation(s)
- Thomas P Corner
- Chemistry Research Laboratory, Department of Chemistry and the Ineos Oxford Institute for Antimicrobial Research, University of Oxford 12 Mansfield Road OX1 3TA Oxford United Kingdom
| | - Ryan Z R Teo
- Chemistry Research Laboratory, Department of Chemistry and the Ineos Oxford Institute for Antimicrobial Research, University of Oxford 12 Mansfield Road OX1 3TA Oxford United Kingdom
| | - Yue Wu
- State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Drug Design and Optimization and Department of Chemistry, China Pharmaceutical University Nanjing 211198 China
| | - Eidarus Salah
- Chemistry Research Laboratory, Department of Chemistry and the Ineos Oxford Institute for Antimicrobial Research, University of Oxford 12 Mansfield Road OX1 3TA Oxford United Kingdom
| | - Yu Nakashima
- Institute of Natural Medicine, University of Toyama 2630-Sugitani 930-0194 Toyama Japan
| | - Giorgia Fiorini
- Chemistry Research Laboratory, Department of Chemistry and the Ineos Oxford Institute for Antimicrobial Research, University of Oxford 12 Mansfield Road OX1 3TA Oxford United Kingdom
| | - Anthony Tumber
- Chemistry Research Laboratory, Department of Chemistry and the Ineos Oxford Institute for Antimicrobial Research, University of Oxford 12 Mansfield Road OX1 3TA Oxford United Kingdom
| | - Amelia Brasnett
- Chemistry Research Laboratory, Department of Chemistry and the Ineos Oxford Institute for Antimicrobial Research, University of Oxford 12 Mansfield Road OX1 3TA Oxford United Kingdom
| | - James P Holt-Martyn
- Chemistry Research Laboratory, Department of Chemistry and the Ineos Oxford Institute for Antimicrobial Research, University of Oxford 12 Mansfield Road OX1 3TA Oxford United Kingdom
| | - William D Figg
- Chemistry Research Laboratory, Department of Chemistry and the Ineos Oxford Institute for Antimicrobial Research, University of Oxford 12 Mansfield Road OX1 3TA Oxford United Kingdom
| | - Xiaojin Zhang
- State Key Laboratory of Natural Medicines, Jiangsu Key Laboratory of Drug Design and Optimization and Department of Chemistry, China Pharmaceutical University Nanjing 211198 China
| | - Lennart Brewitz
- Chemistry Research Laboratory, Department of Chemistry and the Ineos Oxford Institute for Antimicrobial Research, University of Oxford 12 Mansfield Road OX1 3TA Oxford United Kingdom
| | - Christopher J Schofield
- Chemistry Research Laboratory, Department of Chemistry and the Ineos Oxford Institute for Antimicrobial Research, University of Oxford 12 Mansfield Road OX1 3TA Oxford United Kingdom
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10
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Kato S. Lactoferrin inhibits the proliferation of IMR‑32 neuroblastoma cells even under X‑rays. MEDICINE INTERNATIONAL 2023; 3:33. [PMID: 37448769 PMCID: PMC10336960 DOI: 10.3892/mi.2023.93] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Subscribe] [Scholar Register] [Received: 04/03/2023] [Accepted: 06/19/2023] [Indexed: 07/15/2023]
Abstract
Neuroblastoma is a typical solid tumor common in childhood. The present study investigated the inhibitory effects of lactoferrin on the proliferation of IMR-32 neuroblastoma cells, including under X-ray irradiation. In controlled in vitro assays, it was found that lactoferrin inhibited cell proliferation, accompanied by cell membrane disruption. Furthermore, intracellular reactive oxygen species generation increased in IMR-32 cells treated with lactoferrin, causing membrane lipid peroxidation and the leakage of lactate dehydrogenase. The IC50 values for cell proliferation were ~2.0 nM for doxorubicin, 2.7 mM for dibutyryl-cAMP and 45.9 µM for lactoferrin. X-ray irradiation at 1 Gy decreased cell proliferation to ~30%, which was not restored by lactoferrin. In the Fenton reaction system with iron chloride, lactoferrin increased hydroxyl radical (OH·) formation via H2O2, as confirmed by electron spin resonance spectra. On the whole, the findings of the present study indicate that lactoferrin, found abundantly in milk, may help prevent or treat neuroblastoma in infants with modest efficacy, and does not exert a protective effect against X-rays.
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Affiliation(s)
- Shinya Kato
- Radioisotope Experimental Facility, Advanced Science Research Promotion Center, Mie University, Tsu, Mie 514-8507, Japan
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11
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Sharon E, Pietrokovski Y, Engel I, Assali R, Houri-Haddad Y, Beyth N. Biocompatibility, Surface Morphology, and Bacterial Load of Dental Implant Abutments following Decontamination Protocols: An In-Vitro Study. MATERIALS (BASEL, SWITZERLAND) 2023; 16:ma16114080. [PMID: 37297212 DOI: 10.3390/ma16114080] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 03/22/2023] [Revised: 05/14/2023] [Accepted: 05/16/2023] [Indexed: 06/12/2023]
Abstract
The long-term success of dental implant rehabilitation depends significantly on proper peri-implant soft tissue integration. Therefore, decontamination of abutments prior to their connection to the implant is beneficial to enhance soft tissue attachment and to aid in maintaining marginal bone around the implant. Consequently, different implant abutment decontamination protocols were evaluated regarding biocompatibility, surface morphology, and bacterial load. The protocols evaluated were autoclave sterilization, ultrasonic washing, steam cleaning, chlorhexidine chemical decontamination, and sodium hypochlorite chemical decontamination. The control groups included: (1) implant abutments prepared and polished in a dental lab without decontamination and (2) unprepared implant abutments obtained directly from the company. Surface analysis was performed using scanning electron microscopy (SEM). Biocompatibility was evaluated using XTT cell viability and proliferation assays. Biofilm biomass and viable counts (CFU/mL) (n = 5 for each test) were used for surface bacterial load evaluation. Surface analysis revealed areas of debris and accumulation of materials, such as iron, cobalt, chromium, and other metals, in all abutments prepared by the lab and with all decontamination protocols. Steam cleaning was the most efficient method for reducing contamination. Chlorhexidine and sodium hypochlorite left residual materials on the abutments. XTT results showed that the chlorhexidine group (M = 0.7005, SD = 0.2995) had the lowest values (p < 0.001) (autoclave: M = 3.6354, SD = 0.1510; ultrasonic: M = 3.4077, SD = 0.3730; steam: M = 3.2903, SD = 0.2172; NaOCl: M = 3.5377, SD = 0.0927; prep non-decont.: M = 3.4815, SD = 0.2326; factory: M = 3.6173, SD = 0.0392). Bacterial growth (CFU/mL) was high in the abutments treated with steam cleaning and ultrasonic bath: 2.93 × 109, SD = 1.68 × 1012 and 1.83 × 109, SD = 3.95 × 1010, respectively. Abutments treated with chlorhexidine showed higher toxicity to cells, while all other samples showed similar effects to the control. In conclusion, steam cleaning seemed to be the most efficient method for reducing debris and metallic contamination. Bacterial load can be reduced using autoclaving, chlorhexidine, and NaOCl.
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Affiliation(s)
- Esi Sharon
- Department of Prosthodontics, Hadassah Medical Center, Faculty of Dental Medicine, Hebrew University of Jerusalem, Jerusalem 9112102, Israel
| | - Yoav Pietrokovski
- Department of Prosthodontics, Hadassah Medical Center, Faculty of Dental Medicine, Hebrew University of Jerusalem, Jerusalem 9112102, Israel
| | - Ilana Engel
- Department of Prosthodontics, Hadassah Medical Center, Faculty of Dental Medicine, Hebrew University of Jerusalem, Jerusalem 9112102, Israel
| | - Rula Assali
- Department of Prosthodontics, Hadassah Medical Center, Faculty of Dental Medicine, Hebrew University of Jerusalem, Jerusalem 9112102, Israel
| | - Yael Houri-Haddad
- Department of Prosthodontics, Hadassah Medical Center, Faculty of Dental Medicine, Hebrew University of Jerusalem, Jerusalem 9112102, Israel
| | - Nurit Beyth
- Department of Prosthodontics, Hadassah Medical Center, Faculty of Dental Medicine, Hebrew University of Jerusalem, Jerusalem 9112102, Israel
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12
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Kim MY, Kim S, Lee J, Kim JI, Oh E, Kim SW, Lee E, Cho KS, Kim CS, Lee MH. Lignan-Rich Sesame ( Sesamum indicum L.) Cultivar Exhibits In Vitro Anti-Cholinesterase Activity, Anti-Neurotoxicity in Amyloid-β Induced SH-SY5Y Cells, and Produces an In Vivo Nootropic Effect in Scopolamine-Induced Memory Impaired Mice. Antioxidants (Basel) 2023; 12:antiox12051110. [PMID: 37237976 DOI: 10.3390/antiox12051110] [Citation(s) in RCA: 4] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/03/2023] [Revised: 04/28/2023] [Accepted: 05/15/2023] [Indexed: 05/28/2023] Open
Abstract
Alzheimer's disease, a major cause of dementia, is characterized by impaired cholinergic function, increased oxidative stress, and amyloid cascade induction. Sesame lignans have attracted considerable attention owing to their beneficial effects on brain health. This study investigated the neuroprotective potential of lignan-rich sesame cultivars. Among the 10 sesame varieties studied, Milyang 74 (M74) extracts exhibited the highest total lignan content (17.71 mg/g) and in vitro acetylcholinesterase (AChE) inhibitory activity (66.17%, 0.4 mg/mL). M74 extracts were the most effective in improving cell viability and inhibiting reactive oxygen species (ROS) and malondialdehyde (MDA) generation in amyloid-β25-35 fragment-treated SH-SY5Y cells. Thus, M74 was used to evaluate the nootropic effects of sesame extracts and oil on scopolamine (2 mg/kg)-induced memory impairment in mice compared to the control cultivar (Goenback). Pretreatment with the M74 extract (250 and 500 mg/kg) and oil (1 and 2 mL/kg) effectively improved memory disorder in mice (demonstrated by the passive avoidance test), inhibited AChE, and enhanced acetylcholine (Ach) levels. Moreover, immunohistochemistry and Western blot results showed that the M74 extract and oil reversed the scopolamine-induced increase in APP, BACE-1, and presenilin expression levels in the amyloid cascade and decreased BDNF and NGF expression levels in neuronal regeneration.
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Affiliation(s)
- Min-Young Kim
- Department of Southern Area Crop Science, National Institute of Crop Science, Rural Development Administration, Milyang 50424, Republic of Korea
| | - Sungup Kim
- Department of Southern Area Crop Science, National Institute of Crop Science, Rural Development Administration, Milyang 50424, Republic of Korea
| | - Jeongeun Lee
- Department of Southern Area Crop Science, National Institute of Crop Science, Rural Development Administration, Milyang 50424, Republic of Korea
| | - Jung-In Kim
- Department of Southern Area Crop Science, National Institute of Crop Science, Rural Development Administration, Milyang 50424, Republic of Korea
| | - Eunyoung Oh
- Department of Southern Area Crop Science, National Institute of Crop Science, Rural Development Administration, Milyang 50424, Republic of Korea
| | - Sang-Woo Kim
- Department of Southern Area Crop Science, National Institute of Crop Science, Rural Development Administration, Milyang 50424, Republic of Korea
| | - Eunsoo Lee
- Department of Southern Area Crop Science, National Institute of Crop Science, Rural Development Administration, Milyang 50424, Republic of Korea
| | - Kwang-Soo Cho
- Department of Southern Area Crop Science, National Institute of Crop Science, Rural Development Administration, Milyang 50424, Republic of Korea
| | - Choon-Song Kim
- Department of Southern Area Crop Science, National Institute of Crop Science, Rural Development Administration, Milyang 50424, Republic of Korea
| | - Myoung-Hee Lee
- Department of Southern Area Crop Science, National Institute of Crop Science, Rural Development Administration, Milyang 50424, Republic of Korea
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13
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Yang CZ, Guo W, Wang YF, Hu LH, Wang J, Luo JM, Yao XH, Liu S, Tao LT, Sun LL, Lin LZ. Reduction in gefitinib resistance mediated by Yi-Fei San-Jie pill in non-small cell lung cancer through regulation of tyrosine metabolism, cell cycle, and the MET/EGFR signaling pathway. JOURNAL OF ETHNOPHARMACOLOGY 2023; 314:116566. [PMID: 37169317 DOI: 10.1016/j.jep.2023.116566] [Citation(s) in RCA: 2] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 02/20/2023] [Revised: 04/16/2023] [Accepted: 04/29/2023] [Indexed: 05/13/2023]
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE The Chinese herbal prescription Yi-Fei San-Jie pill (YFSJ) has been used for adjuvant treatment in patients with lung cancer for a long time. AIM OF THE STUDY Reports have indicated that the combination of gefitinib (Gef) with YFSJ inhibits the proliferation of EGFR-TKI-resistant cell lines by enhancing cellular apoptosis and autophagy in non-small cell lung cancer (NSCLC). However, the molecular mechanisms underlying the effect of YFSJ on EGFR-TKI resistance and related metabolic pathways remain to be explored. MATERIALS AND METHODS In our report, ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), metabolomics, network pharmacology, bioinformatics, and biological analysis methods were used to investigate the mechanism. RESULTS The UPLC-MS/MS data identified 42 active compounds of YFSJ extracts. YFSJ extracts can enhance the antitumor efficacy of Gef without hepatic and renal toxicity in vivo. The analysis of the metabolomics pathway enrichment revealed that YFSJ mainly affected the tyrosine metabolism pathway in rat models. Moreover, YFSJ has been shown to reverse Gef resistance and improve the effects of Gef on the cellular viability, migration capacity, and cell cycle arrest of NSCLC cell lines with EGFR mutations. The results of network pharmacology and molecular docking analyses revealed that tyrosine metabolism-related active compounds of YFSJ affect EGFR-TKIs resistance in NSCLC by targeting cell cycle and the MET/EGFR signaling pathway; these findings were validated by western blotting and immunohistochemistry. CONCLUSIONS YFSJ inhibits NSCLC by inducing cell cycle arrest in the G1/S phase to suppress tumor growth, cell viability, and cell migration through synergistic effects with Gef via the tyrosine metabolic pathway and the EGFR/MET signaling pathway. To summarize, the findings of the current study indicate that YFSJ is a prospective complementary treatment for Gef-resistant NSCLC.
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Affiliation(s)
- Cai-Zhi Yang
- The First School of Medicine, Guangzhou University of Chinese Medicine, Guangzhou, 510405, China.
| | - Wei Guo
- Department of Oncology, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, 510405, China.
| | - Yi-Fan Wang
- The First School of Medicine, Guangzhou University of Chinese Medicine, Guangzhou, 510405, China.
| | - Lei-Hao Hu
- The First School of Medicine, Guangzhou University of Chinese Medicine, Guangzhou, 510405, China.
| | - Jing Wang
- State Key Laboratory of Quality Research in Chinese Medicines, Faculty of Chinese Medicine, Macau University of Science and Technology, Macau, 999078, China.
| | - Jia-Min Luo
- The First School of Medicine, Guangzhou University of Chinese Medicine, Guangzhou, 510405, China.
| | - Xiao-Hui Yao
- Guangzhou University of Chinese Medicine, Guangzhou, 510405, China.
| | - Shan Liu
- The First School of Medicine, Guangzhou University of Chinese Medicine, Guangzhou, 510405, China.
| | - Lan-Ting Tao
- Department of Oncology, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, 510120, China.
| | - Ling-Ling Sun
- Department of Oncology, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, 510405, China.
| | - Li-Zhu Lin
- Department of Oncology, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, 510405, China.
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14
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Alex J, Mathew TV. Surface Modification of Bi 2O 3 Nanoparticles with Biotinylated β-Cyclodextrin as a Biocompatible Therapeutic Agent for Anticancer and Antimicrobial Applications. Molecules 2023; 28:molecules28083604. [PMID: 37110839 PMCID: PMC10142954 DOI: 10.3390/molecules28083604] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/05/2023] [Revised: 04/12/2023] [Accepted: 04/15/2023] [Indexed: 04/29/2023] Open
Abstract
Bismuth oxide nanoparticles with appropriate surface chemistry exhibit many interesting properties that can be utilized in a variety of applications. This paper describes a new route to the surface modification of bismuth oxide nanoparticles (Bi2O3 NPs) using functionalized beta-Cyclodextrin (β-CD) as a biocompatible system. The synthesis of Bi2O3 NP was done using PVA (poly vinyl alcohol) as the reductant and the Steglich esterification procedure for the functionalization of β-CD with biotin. Ultimately, the Bi2O3 NPs are modified using this functionalized β-CD system. The particle size of the synthesized Bi2O3 NPs is found to be in the range of 12-16 nm. The modified biocompatible systems were characterized using different characterization techniques such as Fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-ray powder diffraction (XRD) and Differential Scanning Calorimetric analysis (DSC). Additionally, the antibacterial and anticancerous effects of the surface-modified Bi2O3 NP system were also investigated.
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Affiliation(s)
- Jogy Alex
- Department of Chemistry, St. Thomas College Palai, Arunapuram P.O., Kottayam 686574, Kerala, India
| | - Thomas V Mathew
- Department of Chemistry, St. Thomas College Palai, Arunapuram P.O., Kottayam 686574, Kerala, India
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15
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Cho HY, Ahn S, Cho YS, Seo SK, Kim DH, Shin JG, Lee SJ. CYP2C19 Contributes to THP-1-Cell-Derived M2 Macrophage Polarization by Producing 11,12- and 14,15-Epoxyeicosatrienoic Acid, Agonists of the PPARγ Receptor. Pharmaceuticals (Basel) 2023; 16:ph16040593. [PMID: 37111350 PMCID: PMC10143178 DOI: 10.3390/ph16040593] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/22/2023] [Revised: 04/11/2023] [Accepted: 04/11/2023] [Indexed: 04/29/2023] Open
Abstract
Although the functional roles of M1 and M2 macrophages in the immune response and drug resistance are important, the expression and role of cytochrome P450s (CYPs) in these cells remain largely unknown. Differential expression of the 12 most common CYPs (CYP1A1, 1A2, 1B1, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, 2J2, 3A4, and 3A5) were screened in THP-1-cell-derived M1 and M2 macrophages using reverse transcription PCR. CYP2C19 was highly expressed in THP-1-cell-derived M2 macrophages, but it was negligibly expressed in THP-1-cell-derived M1 macrophages at the mRNA and protein levels as analyzed by reverse transcription quantitative PCR and Western blot, respectively. CYP2C19 enzyme activity was also very high in THP-1-cell-derived M2 compared to M1 macrophages (> 99%, p < 0.01), which was verified using inhibitors of CYP2C19 activity. Endogenous levels of the CYP2C19 metabolites 11,12-epoxyeicosatrienoic acid (11,12-EET) and 14,15-EET were reduced by 40% and 50% in cells treated with the CYP2C19 inhibitor and by 50% and 60% in the culture medium, respectively. Both 11,12-EET and 14,15-EET were identified as PPARγ agonists in an in vitro assay. When THP-1-cell-derived M2 cells were treated with CYP2C19 inhibitors, 11,12- and 14,15-EETs were significantly reduced, and in parallel with the reduction of these CYP2C19 metabolites, the expression of M2 cell marker genes was also significantly decreased (p < 0.01). Therefore, it was suggested that CYP2C19 may contribute to M2 cell polarization by producing PPARγ agonists. Further studies are needed to understand the endogenous role of CYP2C19 in M2 macrophages with respect to immunologic function and cell polarization.
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Affiliation(s)
- Hee Young Cho
- Department of Pharmacology and PharmacoGenomics Research Center, Inje University College of Medicine, Inje University, Busan 47392, Republic of Korea
- Center for Personalized Precision Medicine of Tuberculosis, Inje University College of Medicine, Inje University, Busan 47392, Republic of Korea
| | - Sangzin Ahn
- Department of Pharmacology and PharmacoGenomics Research Center, Inje University College of Medicine, Inje University, Busan 47392, Republic of Korea
- Center for Personalized Precision Medicine of Tuberculosis, Inje University College of Medicine, Inje University, Busan 47392, Republic of Korea
| | - Yong-Soon Cho
- Department of Pharmacology and PharmacoGenomics Research Center, Inje University College of Medicine, Inje University, Busan 47392, Republic of Korea
- Center for Personalized Precision Medicine of Tuberculosis, Inje University College of Medicine, Inje University, Busan 47392, Republic of Korea
| | - Su-Kil Seo
- Center for Personalized Precision Medicine of Tuberculosis, Inje University College of Medicine, Inje University, Busan 47392, Republic of Korea
- Department of Microbiology and Immunology, Inje University College of Medicine, Inje University, Busan 47392, Republic of Korea
| | - Dong Hyun Kim
- Department of Pharmacology and PharmacoGenomics Research Center, Inje University College of Medicine, Inje University, Busan 47392, Republic of Korea
- Center for Personalized Precision Medicine of Tuberculosis, Inje University College of Medicine, Inje University, Busan 47392, Republic of Korea
| | - Jae-Gook Shin
- Department of Pharmacology and PharmacoGenomics Research Center, Inje University College of Medicine, Inje University, Busan 47392, Republic of Korea
- Center for Personalized Precision Medicine of Tuberculosis, Inje University College of Medicine, Inje University, Busan 47392, Republic of Korea
| | - Su-Jun Lee
- Department of Pharmacology and PharmacoGenomics Research Center, Inje University College of Medicine, Inje University, Busan 47392, Republic of Korea
- Center for Personalized Precision Medicine of Tuberculosis, Inje University College of Medicine, Inje University, Busan 47392, Republic of Korea
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16
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Wu CP, Hung CY, Hsieh YJ, Murakami M, Huang YH, Su TY, Hung TH, Yu JS, Wu YS, Ambudkar SV. ABCB1 and ABCG2 Overexpression Mediates Resistance to the Phosphatidylinositol 3-Kinase Inhibitor HS-173 in Cancer Cell Lines. Cells 2023; 12:cells12071056. [PMID: 37048130 PMCID: PMC10093605 DOI: 10.3390/cells12071056] [Citation(s) in RCA: 4] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/28/2023] [Revised: 03/27/2023] [Accepted: 03/29/2023] [Indexed: 04/03/2023] Open
Abstract
Constitutive activation of the phosphoinositide-3-kinase (PI3K)/Akt signaling pathway is crucial for tumor growth and progression. As such, this pathway has been an enticing target for drug discovery. Although HS-173 is a potent PI3K inhibitor that halts cancer cell proliferation via G2/M cell cycle arrest, the resistance mechanisms to HS-173 have not been investigated. In this study, we investigated the susceptibility of HS-173 to efflux mediated by the multidrug efflux transporters ABCB1 and ABCG2, which are two of the most well-known ATP-binding cassette (ABC) transporters associated with the development of cancer multidrug resistance (MDR). We found that the overexpression of ABCB1 or ABCG2 significantly reduced the efficacy of HS-173 in human cancer cells. Our data show that the intracellular accumulation of HS-173 was substantially reduced by ABCB1 and ABCG2, affecting G2/M arrest and apoptosis induced by HS-173. More importantly, the efficacy of HS-173 in multidrug-resistant cancer cells could be recovered by inhibiting the drug-efflux function of ABCB1 and ABCG2. Taken together, our study has demonstrated that HS-173 is a substrate for both ABCB1 and ABCG2, resulting in decreased intracellular concentration of this drug, which may have implications for its clinical use.
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Affiliation(s)
- Chung-Pu Wu
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
- Department of Physiology and Pharmacology, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
- Molecular Medicine Research Center, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
- Department of Obstetrics and Gynecology, Taipei Chang Gung Memorial Hospital, Taipei 10507, Taiwan
| | - Cheng-Yu Hung
- Molecular Medicine Research Center, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
| | - Ya-Ju Hsieh
- Molecular Medicine Research Center, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
| | - Megumi Murakami
- Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD 20892, USA
| | - Yang-Hui Huang
- Department of Obstetrics and Gynecology, Taipei Chang Gung Memorial Hospital, Taipei 10507, Taiwan
| | - Tsung-Yao Su
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
| | - Tai-Ho Hung
- Department of Obstetrics and Gynecology, Taipei Chang Gung Memorial Hospital, Taipei 10507, Taiwan
- Department of Medicine, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
- Department of Obstetrics and Gynecology, Keelung Chang Gung Memorial Hospital, Keelung 20401, Taiwan
| | - Jau-Song Yu
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
- Molecular Medicine Research Center, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
- Department of Biochemistry and Molecular Biology, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
- Liver Research Center, Linkou Chang Gung Memorial Hospital, Taoyuan 33302, Taiwan
| | - Yu-Shan Wu
- Department of Chemistry, Tunghai University, Taichung 40704, Taiwan
| | - Suresh V. Ambudkar
- Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD 20892, USA
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17
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Zapol’skii VA, Kaul S, Karge B, Brönstrup M, Gjikaj M, Kaufmann DE. A New Way to 2,3,4-Trisubstituted Benzo[h]quinolines: Synthesis, Consecutive Reactions and Cellular Activities †. Molecules 2023; 28:molecules28062479. [PMID: 36985452 PMCID: PMC10058827 DOI: 10.3390/molecules28062479] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/02/2023] [Revised: 02/27/2023] [Accepted: 02/27/2023] [Indexed: 03/11/2023] Open
Abstract
The reaction of mercaptoacetic acid esters with pentachloro-2-nitro-1,3-butadiene provides the appropriate precursors for the synthesis of 2,3,4-trisubstituted benzo[h]quinolines. These heterocycles are easily accessible via a single-step reaction with naphthalen-1-amine or anthracen-1-amine as the precursor. Due to the steric bulk and high electron density ring, the ring closure of benzo[h]quinolines takes place exclusively. Such highly substituted annelated pyridine systems can be modified in subsequent, selective reactions to build up new N-heterocycles with promising microbiological properties. The antibacterial and antiproliferative assays against four mammalian cell lines demonstrate that some of the sulfur-substituted benzo[h]quinoline analogs display potent phenotypic bioactivities in the single-digit micromolar range.
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Affiliation(s)
- Viktor A. Zapol’skii
- Institute of Organic Chemistry, Clausthal University of Technology, Leibnizstraße 6, 38678 Clausthal-Zellerfeld, Germany
| | - Sandra Kaul
- Institute of Organic Chemistry, Clausthal University of Technology, Leibnizstraße 6, 38678 Clausthal-Zellerfeld, Germany
| | - Bianka Karge
- Department of Chemical Biology, Helmholtz Centre for Infection Research, Inhoffenstrasse 7, 38124 Braunschweig, Germany
| | - Mark Brönstrup
- Department of Chemical Biology, Helmholtz Centre for Infection Research, Inhoffenstrasse 7, 38124 Braunschweig, Germany
| | - Mimoza Gjikaj
- Institute of Inorganic and Analytical Chemistry, Clausthal University of Technology, Paul-Ernst-Str. 4, 38678 Clausthal-Zellerfeld, Germany
| | - Dieter E. Kaufmann
- Institute of Organic Chemistry, Clausthal University of Technology, Leibnizstraße 6, 38678 Clausthal-Zellerfeld, Germany
- Correspondence:
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18
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Cotner M, Meng S, Jost T, Gardner A, De Santiago C, Brock A. Integration of quantitative methods and mathematical approaches for the modeling of cancer cell proliferation dynamics. Am J Physiol Cell Physiol 2023; 324:C247-C262. [PMID: 36503241 PMCID: PMC9886359 DOI: 10.1152/ajpcell.00185.2022] [Citation(s) in RCA: 2] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/02/2022] [Revised: 11/21/2022] [Accepted: 11/21/2022] [Indexed: 12/15/2022]
Abstract
Physiological processes rely on the control of cell proliferation, and the dysregulation of these processes underlies various pathological conditions, including cancer. Mathematical modeling can provide new insights into the complex regulation of cell proliferation dynamics. In this review, we first examine quantitative experimental approaches for measuring cell proliferation dynamics in vitro and compare the various types of data that can be obtained in these settings. We then explore the toolbox of common mathematical modeling frameworks that can describe cell behavior, dynamics, and interactions of proliferation. We discuss how these wet-laboratory studies may be integrated with different mathematical modeling approaches to aid the interpretation of the results and to enable the prediction of cell behaviors, specifically in the context of cancer.
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Affiliation(s)
- Michael Cotner
- Department of Biomedical Engineering, The University of Texas at Austin, Austin, Texas
| | - Sarah Meng
- Department of Biomedical Engineering, The University of Texas at Austin, Austin, Texas
| | - Tyler Jost
- Department of Biomedical Engineering, The University of Texas at Austin, Austin, Texas
| | - Andrea Gardner
- Department of Biomedical Engineering, The University of Texas at Austin, Austin, Texas
| | - Carolina De Santiago
- Department of Biomedical Engineering, The University of Texas at Austin, Austin, Texas
| | - Amy Brock
- Department of Biomedical Engineering, The University of Texas at Austin, Austin, Texas
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19
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Design and cytotoxic evaluation via apoptotic and antiproliferative activity for novel 11(4-aminophenylamino)neocryptolepine on hepatocellular and colorectal cancer cells. Apoptosis 2023; 28:653-668. [PMID: 36719468 DOI: 10.1007/s10495-023-01810-y] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Accepted: 01/05/2023] [Indexed: 02/01/2023]
Abstract
The current study evaluated the cytotoxic activity of 11(4-Aminophenylamino)neocryptolepine (APAN), a novel derivative of neocryptolepine, on hepatocellular (HepG2) and colon (HCT-116) carcinoma cell lines as well as, the possible molecular mechanism through which it exerts its cytotoxic activity. The APAN was synthesized and characterized based on their spectral analyses. Scanning for anticancer target of APAN by Swiss software indicated that APAN had highest affinity for protein tyrosine kinase 6 enzyme. Furthermore, Super pred software indicated that APAN can be indicated in hepatic and colorectal cells with 92%. Molecular docking studies indicated that the binding affinity scores of APAN for protein PDB code: 6CZ4 of tyrosine kinase 6 recorded of - 6.6084 and RMSD value of 0.8891°A, while that for protein PDB: 7JL7 of caspase 3 was - 6.1712 and RMSD of 0.8490°A. Treatment of HepG2 and HCT-116 cells with APAN induced cytotoxicity with IC50 of 2.6 and 1.82 μg/mL respectively. In addition, it induced injury and serious morphological changes in cells including, disappearance of microvilli, membrane blebbing, cytoplasmic condensation, and shrunken nucleus with more condensed chromatin. Moreover, APAN significantly increased protein expression of annexin V (apoptotic marker). Furthermore, APAN significantly increased protein expression of caspase 3 and P53. However, it significantly reduced secretion of VEGF protein into the medium and decreased protein expression of PCNA and Ki67 in HepG2 and HCT-116 cells. This study indicated that APAN had cytotoxic activity against HepG2 and HCT-116 cells via increasing the expression of apoptotic proteins and reducing the expression of proliferative proteins.
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20
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Butler P, Pascheto I, Lizzi M, St-Onge R, Lanner C, Guo B, Masilamani T, Pritzker LB, Kovala AT, Parissenti AM. RNA disruption is a widespread phenomenon associated with stress-induced cell death in tumour cells. Sci Rep 2023; 13:1711. [PMID: 36720913 PMCID: PMC9889758 DOI: 10.1038/s41598-023-28635-8] [Citation(s) in RCA: 4] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/09/2022] [Accepted: 01/23/2023] [Indexed: 02/01/2023] Open
Abstract
We have previously shown that neoadjuvant chemotherapy can induce the degradation of tumour ribosomal RNA (rRNA) in patients with advanced breast cancer, a phenomenon we termed "RNA disruption". Extensive tumour RNA disruption during chemotherapy was associated with a post-treatment pathological complete response and improved disease-free survival. The RNA disruption assay (RDA), which quantifies this phenomenon, is now being evaluated for its clinical utility in a large multinational clinical trial. However, it remains unclear if RNA disruption (i) is manifested across many tumour and non-tumour cell types, (ii) can occur in response to cell stress, and (iii) is associated with tumour cell death. In this study, we show that RNA disruption is induced by several mechanistically distinct chemotherapy agents and report that this phenomenon is observed in response to oxidative stress, endoplasmic reticulum (ER) stress, protein translation inhibition and nutrient/growth factor limitation. We further show that RNA disruption is dose- and time-dependent, and occurs in both tumourigenic and non-tumourigenic cell types. Northern blotting experiments suggest that the rRNA fragments generated during RNA disruption stem (at least in part) from the 28S rRNA. Moreover, we demonstrate that RNA disruption is reproducibly associated with three robust biomarkers of cell death: strongly reduced cell numbers, lost cell replicative capacity, and the generation of cells with a subG1 DNA content. Thus, our findings indicate that RNA disruption is a widespread phenomenon exhibited in mammalian cells under stress, and that high RNA disruption is associated with the onset of cell death.
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Affiliation(s)
- Phillipe Butler
- Graduate Program in Chemical Sciences, Laurentian University, Sudbury, ON, Canada
| | - Isabella Pascheto
- Graduate Program in Chemical Sciences, Laurentian University, Sudbury, ON, Canada
| | - Michayla Lizzi
- Graduate Program in Chemical Sciences, Laurentian University, Sudbury, ON, Canada
| | - Renée St-Onge
- Rna Diagnostics, Inc., Sudbury, ON, Canada.,Rna Diagnostics, Inc., Toronto, ON, Canada
| | - Carita Lanner
- Graduate Program in Chemical Sciences, Laurentian University, Sudbury, ON, Canada.,Division of Medical Sciences, Northern Ontario School of Medicine, Sudbury, ON, Canada
| | - Baoqing Guo
- Health Sciences North Research Institute, Sudbury, ON, Canada
| | - Twinkle Masilamani
- Rna Diagnostics, Inc., Sudbury, ON, Canada.,Rna Diagnostics, Inc., Toronto, ON, Canada
| | - Laura B Pritzker
- Rna Diagnostics, Inc., Sudbury, ON, Canada.,Rna Diagnostics, Inc., Toronto, ON, Canada
| | - A Thomas Kovala
- Graduate Program in Chemical Sciences, Laurentian University, Sudbury, ON, Canada.,Division of Medical Sciences, Northern Ontario School of Medicine, Sudbury, ON, Canada
| | - Amadeo M Parissenti
- Graduate Program in Chemical Sciences, Laurentian University, Sudbury, ON, Canada. .,Rna Diagnostics, Inc., Sudbury, ON, Canada. .,Rna Diagnostics, Inc., Toronto, ON, Canada. .,Health Sciences North Research Institute, Sudbury, ON, Canada. .,Division of Medical Sciences, Northern Ontario School of Medicine, Sudbury, ON, Canada.
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21
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Zipperer A, Scheurer J, Kretschmer D. Cytotoxicity Assays as Predictors of the Safety and Efficacy of Antimicrobial Agents. Methods Mol Biol 2023; 2601:153-167. [PMID: 36445583 DOI: 10.1007/978-1-0716-2855-3_8] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 06/16/2023]
Abstract
The development of safe antimicrobial agents is important for the effective treatment of pathogens. From a multitude of discovered inhibitory compounds, only a few antimicrobial agents are able to enter the market. Many antimicrobials are, on the one hand, quite effective in killing pathogens but, on the other hand, cytotoxic to eukaryotic cells. Cell health can be monitored by various methods. Plasma membrane integrity, DNA synthesis, enzyme activity, and reducing conditions within the cell are known indicators of cell viability and cell death. For a comprehensive overview, methods to analyze cytotoxic and hemolytic effects, e.g., lactate dehydrogenase release, cell proliferation analysis, cell viability analysis based on the activity of different intracellular enzymes, and hemolysis assay of antimicrobial compounds on human cells, are described in this updated chapter.
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Affiliation(s)
- Alexander Zipperer
- Infection Biology, Interfaculty Institute for Microbiology and Infection Medicine Tübingen (IMIT), University of Tübingen, Tübingen, Germany
| | - Jasmin Scheurer
- Department of Dermatology, Division of Dermatooncology, University of Tübingen, Tübingen, Germany
| | - Dorothee Kretschmer
- Infection Biology, Interfaculty Institute for Microbiology and Infection Medicine Tübingen (IMIT), University of Tübingen, Tübingen, Germany.
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22
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Zhong J, Yu W, Tang Y, Zhou X. Synchrotron Radiation FTIR Microspectroscopy Study of Biomolecular Alterations in Vincristine-Treated WRL68 Cells at the Single-Cell Level. ACS OMEGA 2022; 7:47274-47284. [PMID: 36570260 PMCID: PMC9773350 DOI: 10.1021/acsomega.2c06622] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 10/14/2022] [Accepted: 11/25/2022] [Indexed: 06/17/2023]
Abstract
The toxic effect of vincristine on hepatocytes has rarely been studied. Synchrotron radiation-based Fourier transform infrared (SR-FTIR) microspectroscopy is a novel technique for investigating drug-cell interaction systems. In this research, the biomolecular alterations in WRL68 cells induced by vincristine treatment were investigated by SR-FTIR microspectroscopy and were further analyzed by multivariate statistical analysis and semiquantitative methods, including principal component analysis (PCA), orthogonal partial least square-discriminant analysis (OPLS-DA), and the peak area ratios of several characteristic IR bands. In vincristine-treated WRL68 cells, alterations in lipid structures and the presence of more long-chain fatty acids were found. A decrease in protein α-helical content relative to β-sheet structures in vincristine-treated WRL68 cells was identified. The nucleic acid content was decreased relative to that of lipids and proteins in WRL68 cells treated with vincristine. These results provide important information about the toxic effect of vincristine on normal liver cells. This research also provides a new approach to reveal the biomolecular alterations in drug-treated hepatocytes by combining SR-FTIR with multivariate statistical analysis and semiquantitative methods.
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23
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Saqa NA, Khalil-Moghaddam S, Shahvelayati AS. DABCO-based ionic liquid-promoted synthesis of indeno-benzofurans derivatives: Investigation of antioxidant and antidiabetic activities. HETEROCYCL COMMUN 2022. [DOI: 10.1515/hc-2022-0153] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022] Open
Abstract
Abstract
A simple and effective method for synthesis of indeno-benzofurans derivatives using polyphenols and ninhydrins is explored using an acidic catalyst based on DABCO (1,4-diaza bicycle [2.2.2] octane)-based ionic liquid. The products of these types of reactions have very low yields without catalysts, but with DABCO-AIL, the yields are excellent, reaction times are reduced, and the media are cleaner. Using infrared (IR), proton nuclear magnetic resonance (1H NMR), Carbon-13 nuclear magnetic resonance (13C NMR), and mass spectrometry, the structures of products can be confirmed. There is evidence that oxidative stress plays a role in the pathophysiology of numerous diseases, including diabetes. Therapeutic antioxidants are promising candidates for the prevention and treatment of such diseases. To investigate the antioxidant properties of all synthesized derivatives, the 2,2-diphenyl-1-picrylhydrazylhydrazyl-hydrate (DPPH) assay was used. Derivatives 3d and 4 with the highest antioxidant effect (with IC50 value of 0.015 µmol/mL) were selected to evaluate the anti-diabetic effect using the Bernfeld method. The best result was seen at 0.8 mg/mL of 4 derivative and results of the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test revealed that 4 at this concentration lacked cellular toxicity, too.
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Affiliation(s)
- Narges Alipour Saqa
- Department of Biology, Yadegar-e-Imam Khomeini (RAH) shahr-e-Rey Branch, Islamic Azad University , Tehran , Iran
| | - Shiva Khalil-Moghaddam
- Department of Biology, Yadegar-e-Imam Khomeini (RAH) shahr-e-Rey Branch, Islamic Azad University , Tehran , Iran
| | - Ashraf Sadat Shahvelayati
- Department of Chemistry, Yadegar-e-Imam Khomeini (RAH) shahr-e-Rey Branch, Islamic Azad University , Tehran , Iran
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24
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Comparative chemical properties, bioactivity, and cytotoxicity of resin-modified calcium silicate-based pulp capping materials on human dental pulp stem cells. Clin Oral Investig 2022; 26:6839-6853. [PMID: 36104606 DOI: 10.1007/s00784-022-04713-5] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/30/2022] [Accepted: 09/07/2022] [Indexed: 12/31/2022]
Abstract
OBJECTIVES This study investigated the cytotoxicity, the residual monomer release, degree of conversion (DC), calcium ion (Ca2+) release, and crystal structure of TheraCal PT (ThPT) by comparison with TheraCal LC (ThLC) and mineral trioxide aggregate (MTA). MATERIALS AND METHODS The cytotoxicity of the cured materials was evaluated on human dental pulp stem cells (hDPSCs) isolated from third molars by the water-soluble tetrazolium salt (WST-1) method. The monomer release and DC of the resin-containing materials were analyzed by high-performance liquid chromatography (HPLC) and Fourier transform infrared (FTIR), respectively. The chemical composition and Ca2+ release of the materials were determined by scanning electronic microscopy-energy-dispersive spectroscopy (SEM-EDS), X-ray diffractometry (XRD), and inductively coupled plasma-optical emission spectroscopy (ICP-OES), respectively. Statistical differences were evaluated with one-way ANOVA, repeated measure ANOVA, and the Tukey test (p < 0.05). RESULTS MTA showed significantly lower cytotoxicity than either ThLC or ThPT after 1, 3, and 7 days (p < 0.05). TEGDMA release of ThPT is significantly higher than ThLC (p < 0.05). All materials showed calcium Ca2+ release, with MTA significantly higher than the others (p < 0.05). CONCLUSIONS MTA showed low cytotoxicity and high Ca2+ release compared to ThLC and ThPT. CLINICAL RELEVANCE The cytotoxicity and residual monomer release of ThLC and ThPT may raise concerns about the viability of hDPSCs. Further investigations with the use of in vivo research models are required to validate in vitro bioactivity properties and the potential adverse biological effects of ThLC and ThPT on hDPSCs.
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25
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Hong CY, Jo YJ, Kim MY, Chung MN, Choi E, Kim Y, Lee J, Jeong HS. Biological activities of sweet potato ( Ipomoea batatas L.) tips and tubers. Food Sci Nutr 2022; 10:4041-4048. [PMID: 36348769 PMCID: PMC9632182 DOI: 10.1002/fsn3.2999] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/07/2021] [Revised: 06/16/2022] [Accepted: 06/17/2022] [Indexed: 03/27/2024] Open
Abstract
This study was conducted to evaluate the biological activities of sweet potato tips and tubers. Antioxidant activity of 2,2-azino-bis 93-ethlbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities had the highest value of 32.45 mg, AAE/g, and 15.10 mg AAE/g, respectively, in 'Pungwonmi' tips. Angiotensin converting enzyme I inhibitory activity ranged between 47.72% in 'Sinjami' tubers and 62.25% in 'Pungwonmi' tips. α-Glucosidase inhibitory activity had the highest value of 78.81% and 62.93% in 'Pungwonmi' tips and 'Juhwangmi' tubers, respectively. In particular, 'Pungwonmi' tips had the most effective inhibiting effect on intracellular reactive oxygen species levels in HepG2 cells. Wound healing assay result revealed that 'Sinjami' showed 75% wound healing effect. For skin whitening, 'Pungwonmi' tips showed 63% activity at 10 mg/ml. These results suggest that sweet potato tips and tubers can be used to develop functional food and cosmetic materials.
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Affiliation(s)
- Chae Young Hong
- Department of Food Science and BiotechnologyChungbuk National UniversityCheongjuKorea
| | - Yeon Jae Jo
- Department of Food Science and BiotechnologyChungbuk National UniversityCheongjuKorea
| | - Min Young Kim
- National Institute of Crop ScienceRural Development AdministrationMiryangKorea
| | - Mi Nam Chung
- Bioenergy Crop Research InstituteRural Development AdministrationMuanKorea
| | - Ehn‐Kyoung Choi
- College of Veterinary MedicineChungbuk National UniversityCheongjuKorea
| | - Yun‐Bae Kim
- College of Veterinary MedicineChungbuk National UniversityCheongjuKorea
| | - Junsoo Lee
- Department of Food Science and BiotechnologyChungbuk National UniversityCheongjuKorea
| | - Heon Sang Jeong
- Department of Food Science and BiotechnologyChungbuk National UniversityCheongjuKorea
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26
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Li YQ, Murakami M, Huang YH, Hung TH, Wang SP, Wu YS, Ambudkar SV, Wu CP. Hydroxygenkwanin Improves the Efficacy of Cytotoxic Drugs in ABCG2-Overexpressing Multidrug-Resistant Cancer Cells. Int J Mol Sci 2022; 23:ijms232112763. [PMID: 36361555 PMCID: PMC9658017 DOI: 10.3390/ijms232112763] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/03/2022] [Revised: 10/18/2022] [Accepted: 10/20/2022] [Indexed: 11/07/2022] Open
Abstract
Hydroxygenkwanin, a flavonoid isolated from the leaves of the Daphne genkwa plant, is known to have pharmacological properties; however, its modulatory effect on multidrug resistance, which is (MDR) mediated by ATP-binding cassette (ABC) drug transporters, has not been investigated. In this study, we examine the interaction between hydroxygenkwanin, ABCB1, and ABCG2, which are two of the most well-characterized ABC transporters known to contribute to clinical MDR in cancer patients. Hydroxygenkwanin is not an efflux substrate of either ABCB1 or ABCG2. We discovered that, in a concentration-dependent manner, hydroxygenkwanin significantly reverses ABCG2-mediated resistance to multiple cytotoxic anticancer drugs in ABCG2-overexpressing multidrug-resistant cancer cells. Although it inhibited the drug transport function of ABCG2, it had no significant effect on the protein expression of this transporter in cancer cells. Experimental data showing that hydroxygenkwanin stimulates the ATPase activity of ABCG2, and in silico docking analysis of hydroxygenkwanin binding to the inward-open conformation of human ABCG2, further indicate that hydroxygenkwanin sensitizes ABCG2-overexpressing cancer cells by binding to the substrate-binding pocket of ABCG2 and attenuating the transport function of ABCG2. This study demonstrates the potential use of hydroxygenkwanin as an effective inhibitor of ABCG2 in drug combination therapy trials for patients with tumors expressing higher levels of ABCG2.
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Affiliation(s)
- Yan-Qing Li
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
| | - Megumi Murakami
- Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD 20892, USA
| | - Yang-Hui Huang
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
- Department of Physiology and Pharmacology, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
| | - Tai-Ho Hung
- Department of Obstetrics and Gynecology, Taipei Chang Gung Memorial Hospital, Taipei 10507, Taiwan
- Department of Medicine, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
- Department of Obstetrics and Gynecology, Keelung Chang Gung Memorial Hospital, Keelung 20401, Taiwan
| | - Shun-Ping Wang
- Department of Orthopedics, Taichung Veterans General Hospital, Taichung 40704, Taiwan
| | - Yu-Shan Wu
- Department of Chemistry, Tunghai University, Taichung 40704, Taiwan
| | - Suresh V. Ambudkar
- Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD 20892, USA
- Correspondence: (S.V.A.); (C.-P.W.); Tel.: +1-240-760-7192 (S.V.A.); +886-3-2118800 (C.-P.W.)
| | - Chung-Pu Wu
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
- Department of Physiology and Pharmacology, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
- Department of Obstetrics and Gynecology, Taipei Chang Gung Memorial Hospital, Taipei 10507, Taiwan
- Correspondence: (S.V.A.); (C.-P.W.); Tel.: +1-240-760-7192 (S.V.A.); +886-3-2118800 (C.-P.W.)
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27
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Wu CP, Hsieh YJ, Tseng HY, Huang YH, Li YQ, Hung TH, Wang SP, Wu YS. The WD repeat-containing protein 5 (WDR5) antagonist WDR5-0103 restores the efficacy of cytotoxic drugs in multidrug-resistant cancer cells overexpressing ABCB1 or ABCG2. Biomed Pharmacother 2022; 154:113663. [PMID: 36081287 DOI: 10.1016/j.biopha.2022.113663] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/19/2022] [Revised: 08/31/2022] [Accepted: 09/02/2022] [Indexed: 11/02/2022] Open
Abstract
The development of multidrug resistance (MDR) is one of the major challenges in the treatment of cancer which is caused by the overexpression of the ATP-binding cassette (ABC) transporters ABCB1 (P-glycoprotein) and/or ABCG2 (BCRP/MXR/ABCP) in cancer cells. These transporters are capable of reducing the efficacy of cytotoxic drugs by actively effluxing them out of cancer cells. Since there is currently no approved treatment for patients with multidrug-resistant tumors, the drug repurposing approach provides an alternative route to identify agents to reverse MDR mediated by ABCB1 and/or ABCG2 in multidrug-resistant cancer cells. WDR5-0103 is a histone H3 lysine 4 (H3K4) methyltransferase inhibitor that disrupts the interaction between the WD repeat-containing protein 5 (WDR5) and mixed-lineage leukemia (MLL) protein. In this study, the effect of WDR5-0103 on MDR mediated by ABCB1 and ABCG2 was determined. We found that in a concentration-dependent manner, WDR5-0103 could sensitize ABCB1- and ABCG2-overexpressing multidrug-resistant cancer cells to conventional cytotoxic drugs. Our results showed that WDR5-0103 reverses MDR and improves drug-induced apoptosis in multidrug-resistant cancer cells by inhibiting the drug-efflux function of ABCB1 and ABCG2, without altering the protein expression of ABCB1 or ABCG2. The potential sites of interactions of WDR5-0103 with the drug-binding pockets of ABCB1 and ABCG2 were predicted by molecular docking. In conclusion, the MDR reversal activity of WDR5-0103 demonstrated here indicates that it could be used in combination therapy to provide benefits to a subset of patients with tumor expressing high levels of ABCB1 or ABCG2.
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Affiliation(s)
- Chung-Pu Wu
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan; Molecular and Medicine Research Center, Chang Gung University, Taoyuan, Taiwan; Department of Physiology and Pharmacology, College of Medicine, Chang Gung University, Taoyuan, Taiwan; Department of Obstetrics and Gynecology, Taipei Chang Gung Memorial Hospital, Taipei, Taiwan.
| | - Ya-Ju Hsieh
- Molecular and Medicine Research Center, Chang Gung University, Taoyuan, Taiwan.
| | - Han-Yu Tseng
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
| | - Yang-Hui Huang
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
| | - Yan-Qing Li
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
| | - Tai-Ho Hung
- Department of Obstetrics and Gynecology, Taipei Chang Gung Memorial Hospital, Taipei, Taiwan; Department of Medicine, College of Medicine, Chang Gung University, Taoyuan, Taiwan; Department of Obstetrics and Gynecology, Keelung Chang Gung Memorial Hospital, Keelung, Taiwan.
| | - Shun-Ping Wang
- Department of Orthopedics, Taichung Veterans General Hospital, Taichung, Taiwan.
| | - Yu-Shan Wu
- Department of Chemistry, Tunghai University, Taichung, Taiwan.
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Synthesis and Characterization of Silver Nanoparticles from Rhizophora apiculata and Studies on Their Wound Healing, Antioxidant, Anti-Inflammatory, and Cytotoxic Activity. Molecules 2022; 27:molecules27196306. [PMID: 36234841 PMCID: PMC9571849 DOI: 10.3390/molecules27196306] [Citation(s) in RCA: 18] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/20/2022] [Revised: 09/17/2022] [Accepted: 09/19/2022] [Indexed: 11/24/2022] Open
Abstract
Silver nanoparticles (AgNPs) have recently gained interest in the medical field because of their biological features. The present study aimed at screening Rhizophora apiculata secondary metabolites, quantifying their flavonoids and total phenolics content, green synthesis and characterization of R. apiculata silver nanoparticles. In addition, an assessment of in vitro cytotoxic, antioxidant, anti-inflammatory and wound healing activity of R. apiculata and its synthesized AgNPs was carried out. The powdered plant material (leaves) was subjected to Soxhlet extraction to obtain R. apiculata aqueous extract. The R. apiculata extract was used as a reducing agent in synthesizing AgNPs from silver nitrate. The synthesized AgNPs were characterized by UV-Vis, SEM-EDX, XRD, FTIR, particle size analyzer and zeta potential. Further aqueous leaf extract of R. apiculata and AgNPs was subjected for in vitro antioxidant, anti-inflammatory, wound healing and cytotoxic activity against A375 (Skin cancer), A549 (Lung cancer), and KB-3-1 (Oral cancer) cell lines. All experiments were repeated three times (n = 3), and the results were given as the mean ± SEM. The flavonoids and total phenolics content in R. apiculata extract were 44.18 ± 0.086 mg/g of quercetin and 53.24 ± 0.028 mg/g of gallic acid, respectively. SEM analysis revealed R. apiculata AgNPs with diameters ranging from 35 to 100 nm. XRD confirmed that the synthesized silver nanoparticles were crystalline in nature. The cytotoxicity cell viability assay revealed that the AgNPs were less toxic (IC50 105.5 µg/mL) compared to the R. apiculata extract (IC50 47.47 µg/mL) against the non-cancerous fibroblast L929 cell line. Antioxidant, anti-inflammatory, and cytotoxicity tests revealed that AgNPs had significantly more activity than the plant extract. The AgNPs inhibited protein denaturation by a mean percentage of 71.65%, which was equivalent to the standard anti-inflammatory medication diclofenac (94.24%). The AgNPs showed considerable cytotoxic effect, and the percentage of cell viability against skin cancer, lung cancer, and oral cancer cell lines was 31.84%, 56.09% and 22.59%, respectively. R. apiculata AgNPs demonstrated stronger cell migration and percentage of wound closure (82.79%) compared to the plant extract (75.23%). The overall results revealed that R. apiculata AgNPs exhibited potential antioxidant, anti-inflammatory, wound healing, and cytotoxic properties. In future, R. apiculata should be further explored to unmask its therapeutic potential and the mechanistic pathways of AgNPs should be studied in detail in in vivo animal models.
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29
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Addressing artifacts of colorimetric anticancer assays for plant-based drug development. MEDICAL ONCOLOGY (NORTHWOOD, LONDON, ENGLAND) 2022; 39:198. [PMID: 36071299 DOI: 10.1007/s12032-022-01791-z] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 06/04/2022] [Accepted: 07/06/2022] [Indexed: 10/14/2022]
Abstract
Cancer has become the silent killer in less-developed countries and the most significant cause of morbidity worldwide. The accessible and frequently used treatments include surgery, radiotherapy, chemotherapy, and immunotherapy. Chemotherapeutic drugs traditionally involve using plant-based medications either in the form of isolated compounds or as scaffolds for synthetic drugs. To launch a drug in the market, it has to pass through several intricate steps. The multidrug resistance in cancers calls for novel drug discovery and development. Every year anticancer potential of several plant-based compounds and extracts is reported but only a few advances to clinical trials. The false-positive or negative results impact the progress of the cell-based anticancer assays. There are several cell-based assays but the widely used include MTT, MTS, and XTT. In this article, we have discussed various pitfalls and workable solutions.
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30
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A Novel Class of Potent Anti-Tyrosinase Compounds with Antioxidant Activity, 2-(Substituted phenyl)-5-(trifluoromethyl)benzo[ d]thiazoles: In Vitro and In Silico Insights. Antioxidants (Basel) 2022; 11:antiox11071375. [PMID: 35883866 PMCID: PMC9311798 DOI: 10.3390/antiox11071375] [Citation(s) in RCA: 9] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/08/2022] [Revised: 07/10/2022] [Accepted: 07/12/2022] [Indexed: 11/25/2022] Open
Abstract
Sixteen compounds bearing a benzothiazole moiety were synthesized as potential tyrosinase inhibitors and evaluated for mushroom tyrosinase inhibitory activity. The compound 4-(5-(trifluoromethyl)benzo[d]thiazol-2-yl)benzene-1,3-diol (compound 1b) exhibited the highest tyrosinase activity inhibition, with an IC50 value of 0.2 ± 0.01 μM (a potency 55-fold greater than kojic acid). In silico results using mushroom tyrosinase and human tyrosinase showed that the 2,4-hydroxyl substituents on the phenyl ring of 1b played an important role in the inhibition of both tyrosinases. Kinetic studies on mushroom tyrosinase indicated that 1b is a competitive inhibitor of monophenolase and diphenolase, and this was supported by docking results. In B16F10 murine melanoma cells, 1a and 1b dose-dependently and significantly inhibited melanin production intracellularly, and melanin release into medium more strongly than kojic acid, and these effects were attributed to the inhibition of cellular tyrosinase. Furthermore, the inhibition of melanin production by 1b was found to be partially due to the inhibition of tyrosinase glycosylation and the suppression of melanogenesis-associated genes. Compound 1c, which has a catechol group, exhibited potent antioxidant activities against ROS, DPPH, and ABTS, and 1b also had strong ROS and ABTS radical scavenging activities. These results suggest that 5-(trifluoromethyl)benzothiazole derivatives are promising anti-tyrosinase lead compounds with potent antioxidant effects.
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31
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Mahmoudi A, Kesharwani P, Majeed M, Teng Y, Sahebkar A. Recent advances in nanogold as a promising nanocarrier for curcumin delivery. Colloids Surf B Biointerfaces 2022; 215:112481. [PMID: 35453063 DOI: 10.1016/j.colsurfb.2022.112481] [Citation(s) in RCA: 31] [Impact Index Per Article: 10.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/29/2021] [Revised: 03/18/2022] [Accepted: 03/23/2022] [Indexed: 12/12/2022]
Abstract
Curcumin is a natural polyphenolic compound that has promising therapeutic benefits. However, curcumin suffers from low aqueous solubility and poor bioavailability following oral administration, which are severe constraints to its full therapeutic potential. An exciting approach to resolving such challenges has been to incorporate curcumin into gold nanoparticles (AuNPs) to improve its unfavorable physicochemical and biopharmaceutical properties. Growing evidence shows that AuNPs increase cytotoxicity and apoptotic effect of curcumin on cancer cells. Moreover, AuNPs has the potential to enhance curcumin's cellular uptake and antioxidant properties. In addition, numerous benefits have been suggested for exploiting the curcumin's gold (Au) NPs as simple preparation and functionalization. Therefore, we can take advantage of the nanogold combination with curcumin in several therapeutic methods like photothermal therapy and theranostic nanocarrier. Here, we focus on the therapeutic properties of Au/curcumin NPs and the way to improve biocompatibility and bioavailability for curcumin encapsulation, intending to enhance their anticancer and antioxidant capacities. The present review also discusses the utilization and impact of Au NPs as a drug/gene delivery system/platform and various methods for the synthesis of Au/curcumin NPs.
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Affiliation(s)
- Ali Mahmoudi
- Student Research Committee, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran; Department of Medical Biotechnology and Nanotechnology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
| | - Prashant Kesharwani
- Department of Pharmaceutics, School of Pharmaceutical Education and Research, Jamia Hamdard, New Delhi 110062, India.
| | | | - Yong Teng
- Department of Hematology and Medical Oncology, Winship Cancer Institute, Emory University School of Medicine, Atlanta, GA 30322, USA
| | - Amirhossein Sahebkar
- Applied Biomedical Research Center, Mashhad University of Medical Sciences, Mashhad, Iran; Biotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran; School of Medicine, The University of Western Australia, Perth, Australia; Department of Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
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Wu CP, Murakami M, Wu YS, Lin CL, Li YQ, Huang YH, Hung TH, Ambudkar SV. The multi-targeted tyrosine kinase inhibitor SKLB610 resensitizes ABCG2-overexpressing multidrug-resistant cancer cells to chemotherapeutic drugs. Biomed Pharmacother 2022; 149:112922. [PMID: 36068781 PMCID: PMC10506422 DOI: 10.1016/j.biopha.2022.112922] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/17/2022] [Revised: 03/30/2022] [Accepted: 04/01/2022] [Indexed: 11/18/2022] Open
Abstract
The overexpression of ATP-binding cassette (ABC) transporter ABCB1 (P-glycoprotein) or ABCG2 (BCRP/MXR/ABCP) in cancer cells is frequently associated with the development of multidrug resistance (MDR) in cancer patients, which remains a major obstacle to effective cancer treatment. By utilizing energy derived from ATP hydrolysis, both transporters have been shown to reduce the chemosensitivity of cancer cells by actively effluxing cytotoxic anticancer drugs out of cancer cells. Knowing that there are presently no approved drugs or other therapeutics for the treatment of multidrug-resistant cancers, in recent years, studies have investigated the repurposing of tyrosine kinase inhibitors (TKIs) to act as agents against MDR mediated by ABCB1 and/or ABCG2. SKLB610 is a multi-targeted TKI with potent activity against vascular endothelial growth factor receptor 2 (VEGFR2), platelet-derived growth factor receptor (PDGFR), and fibroblast growth factor receptor 2 (FGFR2). In this study, we investigate the interaction of SKLB610 with ABCB1 and ABCG2. We discovered that neither ABCB1 nor ABCG2 confers resistance to SKLB610, but SKLB610 selectively sensitizes ABCG2-overexpressing multidrug-resistant cancer cells to cytotoxic anticancer agents in a concentration-dependent manner. Our data indicate that SKLB610 reverses ABCG2-mediated MDR by attenuating the drug-efflux function of ABCG2 without affecting its total cell expression. These findings are further supported by results of SKLB610-stimulated ABCG2 ATPase activity and in silico docking of SKLB610 in the drug-binding pocket of ABCG2. In summary, we reveal the potential of SKLB610 to overcome resistance to cytotoxic anticancer drugs, which offers an additional treatment option for patients with multidrug-resistant cancers and warrants further investigation.
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Affiliation(s)
- Chung-Pu Wu
- Graduate Institute of Biomedical Sciences, Chang Gung University, Taoyuan 33302, Taiwan; Department of Physiology and Pharmacology, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan; Department of Obstetrics and Gynecology, Taipei Chang Gung Memorial Hospital, Taipei 10507, Taiwan.
| | - Megumi Murakami
- Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD 20892, United States
| | - Yu-Shan Wu
- Department of Chemistry, Tunghai University, Taichung 40704, Taiwan
| | - Chun-Ling Lin
- Graduate Institute of Biomedical Sciences, Chang Gung University, Taoyuan 33302, Taiwan
| | - Yan-Qing Li
- Graduate Institute of Biomedical Sciences, Chang Gung University, Taoyuan 33302, Taiwan
| | - Yang-Hui Huang
- Graduate Institute of Biomedical Sciences, Chang Gung University, Taoyuan 33302, Taiwan; Department of Physiology and Pharmacology, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan
| | - Tai-Ho Hung
- Department of Obstetrics and Gynecology, Taipei Chang Gung Memorial Hospital, Taipei 10507, Taiwan; Department of Medicine, College of Medicine, Chang Gung University, Taoyuan 33302, Taiwan; Department of Obstetrics and Gynecology, Keelung Chang Gung Memorial Hospital, Keelung 20401, Taiwan
| | - Suresh V Ambudkar
- Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD 20892, United States
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Kato S. Under lithium carbonate administration, nicotine triggers cell dysfunction in human glioblastoma U-251MG cells, which is distinct from cotinine. MEDICINE INTERNATIONAL 2022; 2:19. [PMID: 36698501 PMCID: PMC9829207 DOI: 10.3892/mi.2022.44] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 05/03/2022] [Accepted: 05/30/2022] [Indexed: 01/28/2023]
Abstract
Nicotine is an alkaloid found in tobacco leaves. Smoking prevention has been a neglected issue in psychiatry; nicotine intake in conjunction with the administration of the mood stabilizer, lithium carbonate (Li2CO3), may negatively affect brain cells. The present study investigated the combined effects of nicotine and its metabolite, cotinine, and Li2CO3 compared to acetylcholine and dopamine in U-251MG human glioblastoma cells. Cell proliferation was found to be decreased by nicotine and to be further suppressed following treatment with Li2CO3, accompanied by mitotic catastrophe and increased levels of superoxide anion radicals. By contrast, cotinine did not exert such detrimental effects. It was also found that acetylcholine did not suppress cell proliferation, whereas dopamine in conjunction with Li2CO3 decreased cell proliferation in a concentration-dependent manner. The nicotine-induced cell growth inhibition was restored by mecamylamine, a non-competitive antagonist of nicotinic acetylcholine receptors. On the whole, the findings of the present study suggest that nicotine combined with Li2CO3 leads to the suppression of the proliferation of human glioblastoma cells accompanied by mitotic catastrophe and superoxide anion radical generation. These findings may provide further cellular biological insight into the risks associated with smoking under Li2CO3 administration.
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Affiliation(s)
- Shinya Kato
- Radioisotope Experimental Facility, Advanced Science Research Promotion Center, Mie University, Tsu, Mie 514-8507, Japan
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Jo K, Kim S, Yu KW, Chung YB, Kim WJ, Suh HJ, Kim H. Changes in the component sugar and immunostimulating activity of polysaccharides isolated from Dendrobium officinale in the pretreatments. JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE 2022; 102:3021-3028. [PMID: 34775614 DOI: 10.1002/jsfa.11642] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 06/25/2021] [Revised: 08/29/2021] [Accepted: 11/14/2021] [Indexed: 06/13/2023]
Abstract
BACKGROUND To isolate polysaccharides with enhanced immunostimulatory activity from Dendrobium officinale, which is used as a herbal medicine in China and Southeast Asia, D. officinale (DO) was pretreated with organic solvents (DOOS) or puffing at 7.5 and 9.0 kgf (7.5DO and 9DO). Hot-water extracts (DOOS-HW, 7.5DO-HW and 9DO-HW) were prepared from each pretreated DO, along with non-pretreated DO, and crude polysaccharides (DO-CP, DOOS-CP, 7.5DO-CP and 9DO-CP) were fractionated from each hot-water extract using ethanol (five volumes). RESULTS When their immunostimulatory activities were compared by macrophage stimulation and intestinal immune system modulation via Peyer's patches, DOOS-CP showed more potent activity than DO-CP. However, crude polysaccharides fractionated from puffed DO showed significantly lower activity than non-puffed DO and DOOS. The most active polysaccharide contained 95% or more neutral sugar, and the composition ratio of mannose and glucose was 3.0, whereas the lowest polysaccharide content was 2.0 or less. In addition, DOOS-CP was a somewhat refined fraction containing a major peak, representing a molecular weight of 250 kDa, despite being a crude polysaccharide. CONCLUSION These results suggest that pretreatment of D. officinale with organic solvents may enhance the immunostimulatory activity of polysaccharides and affect the mannose/glucose ratio of polysaccharides, which plays an important role in immunostimulation. © 2021 Society of Chemical Industry.
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Affiliation(s)
- Kyungae Jo
- Department of Integrated Biomedical and Life Science, Graduate School, Korea University, Seoul, Republic of Korea
| | - Singeun Kim
- Department of Integrated Biomedical and Life Science, Graduate School, Korea University, Seoul, Republic of Korea
| | - Kwang-Won Yu
- Department of Food and Nutrition, Korea National University of Transportation, Chungbuk, Republic of Korea
| | - Young Bae Chung
- Department of Integrated Biomedical and Life Science, Graduate School, Korea University, Seoul, Republic of Korea
- Research and Development Division, World Institute of Kimchi, Gwangju, Republic of Korea
| | - Woo Jung Kim
- Biocenter, Gyeonggido Business and Science Accelerator, Suwon, Republic of Korea
| | - Hyung Joo Suh
- Department of Integrated Biomedical and Life Science, Graduate School, Korea University, Seoul, Republic of Korea
| | - Hoon Kim
- College of Biotechnology and Natural Resources, Chung-Ang University, Anseong, Republic of Korea
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35
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Hu ZZ, Sha XM, Zhang L, Zha MJ, Tu ZC. From Fish Scale Gelatin to Tyrosinase Inhibitor: A Novel Peptides Screening Approach Application. Front Nutr 2022; 9:853442. [PMID: 35369091 PMCID: PMC8973439 DOI: 10.3389/fnut.2022.853442] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/12/2022] [Accepted: 02/07/2022] [Indexed: 11/13/2022] Open
Abstract
Bioaffinity ultrafiltration combined with LC-Orbitrap-MS/MS was applied for the first time to achieve rapid screening and identification of tyrosinase inhibitory peptides (TYIPs) from grass carp scale gelatin hydrolysates. The binding mode of TYIPs with tyrosinase was investigated by molecular docking technology. The whitening effect of TYIPs was further studied by evaluating the tyrosinase activity and melanin content in mouse B16F10 cells. Four new TYIPs were screened from hydrolysates, among which DLGFLARGF showed the strongest tyrosinase inhibition with an IC50 value of 3.09 mM. Molecular docking showed that hydrogen bonds were the main driving force in the interaction between the peptide DLGFLARGF and tyrosinase. The addition of DLGFLARGF significantly inhibited the tyrosinase activity and melanin production of B16F10 melanoma cells. These results suggest that DLGFLARGF is a promising skin whitening agent for the treatment of potential pigment-related diseases.
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Affiliation(s)
- Zi-Zi Hu
- National R&D Center for Freshwater Fish Processing, College of Chemistry and Chemical Engineering & College of Life Science, Jiangxi Normal University, Nanchang, China
| | - Xiao-Mei Sha
- National R&D Center for Freshwater Fish Processing, College of Chemistry and Chemical Engineering & College of Life Science, Jiangxi Normal University, Nanchang, China
- *Correspondence: Xiao-Mei Sha
| | - Lu Zhang
- National R&D Center for Freshwater Fish Processing, College of Chemistry and Chemical Engineering & College of Life Science, Jiangxi Normal University, Nanchang, China
| | - Min-Jun Zha
- National R&D Center for Freshwater Fish Processing, College of Chemistry and Chemical Engineering & College of Life Science, Jiangxi Normal University, Nanchang, China
| | - Zong-Cai Tu
- National R&D Center for Freshwater Fish Processing, College of Chemistry and Chemical Engineering & College of Life Science, Jiangxi Normal University, Nanchang, China
- State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, China
- Zong-Cai Tu
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Khatua S, Simal-Gandara J, Acharya K. Understanding immune-modulatory efficacy in vitro. Chem Biol Interact 2022; 352:109776. [PMID: 34906553 PMCID: PMC8665649 DOI: 10.1016/j.cbi.2021.109776] [Citation(s) in RCA: 33] [Impact Index Per Article: 11.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/24/2021] [Revised: 11/19/2021] [Accepted: 12/07/2021] [Indexed: 01/07/2023]
Abstract
Boosting or suppressing our immune system represents an attractive adjunct in the treatment of infections including SARS-CoV-2, cancer, AIDS, malnutrition, age related problems and some inflammatory disorders. Thus, there has been a growing interest in exploring and developing novel drugs, natural or synthetic, that can manipulate our defence mechanism. Many of such studies, reported till date, have been designed to explore effect of the therapeutic on function of macrophages, being a key component in innate immune system. Indeed, RAW264.7, J774A.1, THP-1 and U937 cell lines act as ideal model systems for preliminary investigation and selection of dose for in vivo studies. Several bioassays have been standardized so far where many techniques require high throughput instruments, cost effective reagents and technical assistance that may hinder many scholars to perform a method demanding compilation of available protocols. In this review, we have taken an attempt for the first time to congregate commonly used in vitro immune-modulating techniques explaining their principles. The study detected that among about 40 different assays and more than 150 sets of primers, the methods of cell proliferation by MTT, phagocytosis by neutral red, NO detection by Griess reaction and estimation of expression of TLRs, COX-2, iNOS, TNF-α, IL-6 and IL-1β by PCR have been the most widely used to screen the therapeutics under investigation.
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Affiliation(s)
- Somanjana Khatua
- Molecular and Applied Mycology and Plant Pathology Laboratory, Centre of Advanced Study, Department of Botany, University of Calcutta, 35, Ballygunge Circular Road, Kolkata, 700019, West Bengal, India,Department of Botany, Krishnagar Government College, Krishnagar, Nadia, 741101, West Bengal, India
| | - Jesus Simal-Gandara
- Universidade de Vigo, Nutrition and Bromatology Group, Department of Analytical Chemistry and Food Science, Faculty of Science, E-32004, Ourense, Spain,Corresponding author
| | - Krishnendu Acharya
- Molecular and Applied Mycology and Plant Pathology Laboratory, Centre of Advanced Study, Department of Botany, University of Calcutta, 35, Ballygunge Circular Road, Kolkata, 700019, West Bengal, India,Corresponding author
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Al-Qadhi G, Ali Mohammed MM, Al-Ak'hali M, Al-Moraissi EA. Khat (Catha Edulis Forsk) induced apoptosis and cytotoxicity in cultured cells: A scoping review. Heliyon 2021; 7:e08466. [PMID: 34926848 PMCID: PMC8646973 DOI: 10.1016/j.heliyon.2021.e08466] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/01/2021] [Revised: 08/05/2021] [Accepted: 11/19/2021] [Indexed: 11/20/2022] Open
Abstract
BACKGROUND Khat (Catha edulis Forsk) leaves are chewed by people in certain regions of East Africa and the Middle East for their stimulating amphetamine-like effects. The purpose of this scoping review is to systematically map the current in vitro publications that investigated the toxicological potential effects of khat on cultured human or animal cells in terms of cellular viability and activity. METHODS A comprehensive electronic database search was undertaken up to December 2020 without starting date or language restrictions in accordance with the PRISMA extension for scoping review guideline and methodological quality evaluation based on the guidelines for reporting pre-clinical in vitro studies on dental materials. All in vitro studies that investigated the effect of khat plant extract (Catha Edulis) on the cultured human or animal cells were included. RESULTS The initial search yielded 599 articles and 16 articles were finally selected to be included. The treatment of cells with khat produced different degrees of cellular changes, including decreased cellular survival, induction of apoptosis, increased ROS production, alteration of cell phenotype, and of arrest cell cycle. In this contest, khat-exposed cells expressed higher levels of pro-apoptotic protein Bax and lower levels of anti-apoptotic Bcl-2, up-regulated p38, p53, p16, and p21 proteins, as well as premature expression of differentiation markers. CONCLUSION Based on the current scoping review, khat induced apoptosis and cytotoxicity in cultured human cells, including oral cells.
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Affiliation(s)
- Gamilah Al-Qadhi
- Department of Basic Dental Sciences, Faculty of Dentistry, University of Science and Technology, Yemen
| | - Marwan Mansoor Ali Mohammed
- Department of Oral and Craniofacial Health Sciences, College of Dental Medicine, University of Sharjah, United Arab Emirates
| | - Mohammed Al-Ak'hali
- Department of Preventive Dental Sciences, College of Dentistry, Jazan University, Jazan, Saudi Arabia
- Department of Periodontology, Faculty of Dentistry, Sana'a University, Sana'a, Yemen
| | - Essam Ahmed Al-Moraissi
- Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Thamar University, Yemen
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Wu CP, Li YQ, Hung TH, Chang YT, Huang YH, Wu YS. Sophoraflavanone G Resensitizes ABCG2-Overexpressing Multidrug-Resistant Non-Small-Cell Lung Cancer Cells to Chemotherapeutic Drugs. JOURNAL OF NATURAL PRODUCTS 2021; 84:2544-2553. [PMID: 34496204 DOI: 10.1021/acs.jnatprod.1c00584] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/13/2023]
Abstract
Elevated expression of the ATP-binding cassette (ABC) drug transporter ABCG2 in cancer cells contributes to the development of the multidrug resistance phenotype in patients with advanced non-small-cell lung cancer (NSCLC). Due to the lack of U.S. Food and Drug Administration (FDA)-approved synthetic inhibitors of ABCG2, significant efforts have been invested in discovering bioactive compounds of plant origin that are capable of reversing ABCG2-mediated multidrug resistance in cancer cells. Sophoraflavanone G (SFG), a phytoncide isolated from the plant species Sophora flavescens, is known to possess a wide spectrum of pharmacological activities, including antibacterial, anti-inflammatory, antimalarial, and antiproliferative effects. In the present study, the chemosensitizing effect of SFG in ABCG2-overexpressing NSCLC cells was investigated. Experimental results demonstrate that at subtoxic concentrations SFG significantly reversed ABCG2-mediated multidrug resistance in a concentration-dependent manner. Additional biochemical data and in silico docking analysis of SFG to the inward-open conformation of human ABCG2 indicate that SFG inhibited the drug transport function of ABCG2 by interacting with residues within the transmembrane substrate-binding pocket of ABCG2. Collectively, these findings provide evidence that SFG has the potential to be further tested as an effective inhibitor of ABCG2 to improve the efficacy of therapeutic drugs in patients with advanced NSCLC.
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Affiliation(s)
- Chung-Pu Wu
- Department of Obstetrics and Gynecology, Taipei Chang Gung Memorial Hospital, Taipei 33305, Taiwan
| | | | - Tai-Ho Hung
- Department of Obstetrics and Gynecology, Taipei Chang Gung Memorial Hospital, Taipei 33305, Taiwan
| | | | | | - Yu-Shan Wu
- Department of Chemistry, Tunghai University, Taichung 40704, Taiwan
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Zhang Y, Yuan B, Bian B, Zhao H, Kiyomi A, Hayashi H, Iwatani Y, Sugiura M, Takagi N. Cytotoxic Effects of Hellebrigenin and Arenobufagin Against Human Breast Cancer Cells. Front Oncol 2021; 11:711220. [PMID: 34513690 PMCID: PMC8427765 DOI: 10.3389/fonc.2021.711220] [Citation(s) in RCA: 10] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/18/2021] [Accepted: 08/05/2021] [Indexed: 12/12/2022] Open
Abstract
Development of new therapeutic strategies for breast cancer is urgently needed due to the sustained emergence of drug resistance, tumor recurrence and metastasis. To gain a novel insight into therapeutic approaches to fight against breast cancer, the cytocidal effects of hellebrigenin (Helle) and arenobufagin (Areno) were investigated in human estrogen receptor (ER)-positive breast cancer cell line MCF-7 and triple-negative breast cancer cell line MDA-MB-231. Helle exhibited more potent cytotoxicity than Areno in both cancer cells, and MCF-7 cells were more susceptible to both drugs in comparison with MDA-MB-231 cells. Apoptotic-like morphological characteristics, along with the downregulation of the expression level of Bcl-2 and Bcl-xL and the upregulation of the expression level of Bad, were observed in Helle-treated MCF-7 cells. Helle also caused the activation of caspase-8, caspase-9, along with the cleavage of poly(ADP-ribose) polymerase in MCF-7 cells. Helle-mediated necrosis-like phenotype, as evidenced by the increased propidium iodide (PI)-positive cells was further observed. G2/M cell cycle arrest was also induced by Helle in the cells. Upregulation of the expression level of p21 and downregulation of the expression level of cyclin D1, cyclin E1, cdc25C and survivin were observed in MCF-7 cells treated with Helle and occurred in parallel with G2/M arrest. Autophagy was triggered in MCF-7 cells and the addition of wortmannin or 3-MA, two well-known autophagy inhibitors, slightly but significantly rescued the cells. Furthermore, similar alterations of some key molecules associated with the aforementioned biological phenomena were observed in MDA-MB-231 cells. Intriguingly, the numbers of PI-positive cells in Helle-treated MCF-7 cells were significantly reduced by wortmannin and 3-MA, respectively. In addition, Helle-triggered G2/M arrest was significantly corrected by wortmannin, suggesting autophagy induction contributed to Helle-induced cytotoxicity of breast cancer cells by modulating necrosis and cell cycle arrest. Collectively, our results suggested potential usefulness of both Helle and Areno in developing therapeutic strategies to treat patients with different types of breast cancer, especially ER-positive breast cancer.
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Affiliation(s)
- Yu Zhang
- Department of Applied Biochemistry, Tokyo University of Pharmacy & Life Sciences, Hachioji, Japan.,Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, China
| | - Bo Yuan
- Laboratory of Pharmacology, School of Pharmacy, Faculty of Pharmaceutical Sciences, Josai University, Sakado, Japan
| | - Baolin Bian
- Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, China
| | - Haiyu Zhao
- Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing, China
| | - Anna Kiyomi
- Department of Drug Safety and Risk Management, Tokyo University of Pharmacy & Life Sciences, Hachioji, Japan
| | - Hideki Hayashi
- Department of Applied Biochemistry, Tokyo University of Pharmacy & Life Sciences, Hachioji, Japan
| | - Yui Iwatani
- Department of Applied Biochemistry, Tokyo University of Pharmacy & Life Sciences, Hachioji, Japan
| | - Munetoshi Sugiura
- Department of Drug Safety and Risk Management, Tokyo University of Pharmacy & Life Sciences, Hachioji, Japan
| | - Norio Takagi
- Department of Applied Biochemistry, Tokyo University of Pharmacy & Life Sciences, Hachioji, Japan
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Bharadwaz A, Jayasuriya AC. Fabrication of porous chitosan particles using a novel two-step porogen leaching and lyophilization method with the label-free multivariate spectral assessment of live adhered cells. Colloids Surf B Biointerfaces 2021; 208:112094. [PMID: 34500203 DOI: 10.1016/j.colsurfb.2021.112094] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/10/2021] [Revised: 08/27/2021] [Accepted: 08/31/2021] [Indexed: 01/15/2023]
Abstract
Porous chitosan (CS) particles were fabricated using a novel two-step technique that employed a porogen leaching phase followed by lyophilization or freeze-drying. Poly(ethylene glycol) (PEG) was mixed as a porogen in two different quantities with the CS solution before particle synthesis via coacervation. After the PEG leached out into deionized (DI) water at an elevated constant temperature, the final freeze-dried CS particles revealed surface features that resembled pore pockets. A three-dimensional (3D) culture of murine osteoblast cell line (OB-6) was seeded on these particles to analyze the effect of the porous structure on the cell activity, as compared to a control group with no added porogen. The results highlighted an enhancement in cell adhesion and proliferation on the two porous sample groups. A Raman spectroscopy-based label-free technique for live cell biomarker analysis was applied using multivariate spectral analysis. Results of the spectral analysis in the molecular fingerprint region corresponding to the Raman shift between 900 cm-1 and 1700 cm-1inferred inter-group variations. The bands at 1005 cm-1 and 1375 cm-1 were assigned to the live cell biomarkers phenylalanine and glycosaminoglycan, respectively, and were assessed during the multivariate spectral analysis. The corresponding score plot and loading information generated from the Principal Component Analysis (PCA) of the Raman spectrum at day 7 and day 14, pointed at inter-group spectral variations related to cell adhesion and proliferation between the two porous CS particle groups and the control.
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Affiliation(s)
- Angshuman Bharadwaz
- Biomedical Engineering Program, Department of Bioengineering, College of Engineering, The University of Toledo, Toledo, OH, 43606, USA
| | - Ambalangodage C Jayasuriya
- Biomedical Engineering Program, Department of Bioengineering, College of Engineering, The University of Toledo, Toledo, OH, 43606, USA; Department of Orthopaedic Surgery, College of Medicine and Life Sciences, The University of Toledo, Toledo, OH, 43614, USA.
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Wang N, Yang S, Tan T, Huang Y, Chen Y, Dong C, Chen J, Luo X. Tetrandrine suppresses the growth of human osteosarcoma cells by regulating multiple signaling pathways. Bioengineered 2021; 12:5870-5882. [PMID: 34477474 PMCID: PMC8806773 DOI: 10.1080/21655979.2021.1967034] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/02/2022] Open
Abstract
Although osteosarcoma (OS) is the most common malignant tumor among juvenile bone tumors, its treatment plan and clinical outcome have not improved significantly in recent decades. Tetrandrine (TET), a Chinese medicine that is usually used in the therapy of silicosis, hypertension and arthritis, has been confirmed by many studies to possess potent antitumour growth properties, but there are different limitations when describing specific mechanisms. Here, we found that TET can obviously prevent the proliferation, migration and invasion of both 143B and MG63 cells and promote their apoptosis in vitro. Our results for luciferase reporter and Western blotting assays show that TET may exert its antitumour activity by regulating multiplex signaling pathways, including the MAPK/Erk, PTEN/Akt, Juk and Wnt signaling pathways. Furthermore, the regulatory effect of TET on OS cells and related signaling pathways was verified again in vivo. Our findings suggest that the anticancer function of TET on human OS may be mediated by its targeting of multiple signaling pathways and that TET may be used as a single drug or in combination with other drugs during the treatment of OS.
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Affiliation(s)
- Nan Wang
- Department of Orthopedics, The Third Affiliated Hospital of Chongqing Medical University, Chongqing, P.R. China
| | - Shengdong Yang
- Department of Orthopedics, The University-Town Hospital of Chongqing Medical University, Chongqing, P.R. China
| | - Tao Tan
- Department of Orthopedics, The First Affiliated Hospital of Chongqing Medical University, Chongqing, P.R. China
| | - Yanran Huang
- Department of Orthopedics, The First Affiliated Hospital of Chongqing Medical University, Chongqing, P.R. China
| | - Yangmei Chen
- Department of Dermatology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, P.R. China
| | - Chaoqun Dong
- Department of General Surgery, Chongqing Traditional Chinese Medicine Hospital, Chongqing, P.R. China
| | - Jin Chen
- Department of Dermatology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, P.R. China
| | - Xiaoji Luo
- Department of Orthopedics, The First Affiliated Hospital of Chongqing Medical University, Chongqing, P.R. China
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The Second-Generation PIM Kinase Inhibitor TP-3654 Resensitizes ABCG2-Overexpressing Multidrug-Resistant Cancer Cells to Cytotoxic Anticancer Drugs. Int J Mol Sci 2021; 22:ijms22179440. [PMID: 34502348 PMCID: PMC8431370 DOI: 10.3390/ijms22179440] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/14/2021] [Revised: 08/24/2021] [Accepted: 08/27/2021] [Indexed: 12/20/2022] Open
Abstract
Human ATP-binding cassette (ABC) subfamily G member 2 (ABCG2) mediates the transport of a wide variety of conventional cytotoxic anticancer drugs and molecular targeted agents. Consequently, the overexpression of ABCG2 in cancer cells is linked to the development of the multidrug resistance (MDR) phenotype. TP-3654 is an experimental second-generation inhibitor of PIM kinase that is currently under investigation in clinical trials to treat advanced solid tumors and myelofibrosis. In this study, we discovered that by attenuating the drug transport function of ABCG2, TP-3654 resensitizes ABCG2-overexpressing multidrug-resistant cancer cells to cytotoxic ABCG2 substrate drugs topotecan, SN-38 and mitoxantrone. Moreover, our results indicate that ABCG2 does not mediate resistance to TP-3654 and may not play a major role in the induction of resistance to TP-3654 in cancer patients. Taken together, our findings reveal that TP-3654 is a selective, potent modulator of ABCG2 drug efflux function that may offer an additional combination therapy option for the treatment of multidrug-resistant cancers.
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Shamshiddinova M, Gulyamov S, Kim HJ, Jung SH, Baek DJ, Lee YM. A Dansyl-Modified Sphingosine Kinase Inhibitor DPF-543 Enhanced De Novo Ceramide Generation. Int J Mol Sci 2021; 22:ijms22179190. [PMID: 34502095 PMCID: PMC8431253 DOI: 10.3390/ijms22179190] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/03/2021] [Revised: 08/18/2021] [Accepted: 08/19/2021] [Indexed: 12/29/2022] Open
Abstract
Sphingosine-1-phosphate (S1P) synthesized by sphingosine kinase (SPHK) is a signaling molecule, involved in cell proliferation, growth, differentiation, and survival. Indeed, a sharp increase of S1P is linked to a pathological outcome with inflammation, cancer metastasis, or angiogenesis, etc. In this regard, SPHK/S1P axis regulation has been a specific issue in the anticancer strategy to turn accumulated sphingosine (SPN) into cytotoxic ceramides (Cers). For these purposes, there have been numerous chemicals synthesized for SPHK inhibition. In this study, we investigated the comparative efficiency of dansylated PF-543 (DPF-543) on the Cers synthesis along with PF-543. DPF-543 deserved attention in strong cytotoxicity, due to the cytotoxic Cers accumulation by ceramide synthase (CerSs). DPF-543 exhibited dual actions on Cers synthesis by enhancing serine palmitoyltransferase (SPT) activity, and by inhibiting SPHKs, which eventually induced an unusual environment with a high amount of 3-ketosphinganine and sphinganine (SPA). SPA in turn was consumed to synthesize Cers via de novo pathway. Interestingly, PF-543 increased only the SPN level, but not for SPA. In addition, DPF-543 mildly activates acid sphingomyelinase (aSMase), which contributes a partial increase in Cers. Collectively, a dansyl-modified DPF-543 relatively enhanced Cers accumulation via de novo pathway which was not observed in PF-543. Our results demonstrated that the structural modification on SPHK inhibitors is still an attractive anticancer strategy by regulating sphingolipid metabolism.
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Affiliation(s)
- Maftuna Shamshiddinova
- College of Pharmacy, Chungbuk National University, Chungbuk 28160, Korea; (M.S.); (S.G.); (H.-J.K.); (S.-H.J.)
| | - Shokhid Gulyamov
- College of Pharmacy, Chungbuk National University, Chungbuk 28160, Korea; (M.S.); (S.G.); (H.-J.K.); (S.-H.J.)
| | - Hee-Jung Kim
- College of Pharmacy, Chungbuk National University, Chungbuk 28160, Korea; (M.S.); (S.G.); (H.-J.K.); (S.-H.J.)
| | - Seo-Hyeon Jung
- College of Pharmacy, Chungbuk National University, Chungbuk 28160, Korea; (M.S.); (S.G.); (H.-J.K.); (S.-H.J.)
| | - Dong-Jae Baek
- College of Pharmacy, Mokpo National University, Jeonnam 58628, Korea;
| | - Yong-Moon Lee
- College of Pharmacy, Chungbuk National University, Chungbuk 28160, Korea; (M.S.); (S.G.); (H.-J.K.); (S.-H.J.)
- Correspondence: ; Tel.: +82-43-261-2825
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Effects of platinum-coexisting dopamine with X-ray irradiation upon human glioblastoma cell proliferation. Hum Cell 2021; 34:1653-1661. [PMID: 34374034 DOI: 10.1007/s13577-021-00591-3] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/07/2021] [Accepted: 08/02/2021] [Indexed: 10/20/2022]
Abstract
In brain tumors, neurotransmitters and platinum drugs may have some interaction, but their role in radiation therapy remains unclear. We investigated the effects of dopamine in combination with platinum on human glioblastoma U-251MG cells upon X-ray irradiation, comparing with L-DOPA, 2-phenylethylamine and temozolomide. Cell proliferation of U-251MG cells was prominently decreased by dopamine in combination with 10 μM platinum upon 4 Gy of X-ray irradiation, accompanied with intracellular reactive oxygen species generation and mitotic catastrophe. Platinum alone did not increase intracellular reactive oxygen species. On the other hand, L-DOPA in combination with platinum did not decrease cell proliferation regardless of X-ray irradiation. It was clearly shown that 2-phenylethylamine did not suppress cell proliferation as compared to dopamine. Temozolomide decreased cell proliferation in a dose-dependent manner upon X-ray irradiation. However, the combined administration of temozolomide and platinum did not further decrease cell proliferation. The platinum nanoparticles were gradually taken up by cells after administration as determined by ICP analysis. Our results suggest that platinum-coexisting dopamine led cells to mitotic catastrophe due to increased production of intracellular reactive oxygen species which was boosted by X-ray and platinum-catalyzed auto-oxidation of dopamine, and thereby cell proliferation was suppressed. In addition, normal human fibroblast OUMS-36T-1 cells were subjected to experiments. Regarding the effect of the combined administration of dopamine and platinum on each cell which was exposed to X-ray, cell proliferation was decreased in U-251MG cells by the combined administration of platinum, whereas that was not decreased in OUMS-36T-1 cells. This provides one basic insight into the effects of dopamine in combined with platinum on radiation therapy for glioblastoma.
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Mohtar N, Parumasivam T, Gazzali AM, Tan CS, Tan ML, Othman R, Fazalul Rahiman SS, Wahab HA. Advanced Nanoparticle-Based Drug Delivery Systems and Their Cellular Evaluation for Non-Small Cell Lung Cancer Treatment. Cancers (Basel) 2021; 13:3539. [PMID: 34298753 PMCID: PMC8303683 DOI: 10.3390/cancers13143539] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/02/2021] [Revised: 06/18/2021] [Accepted: 06/23/2021] [Indexed: 12/12/2022] Open
Abstract
Lung cancers, the number one cancer killer, can be broadly divided into small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC), with NSCLC being the most commonly diagnosed type. Anticancer agents for NSCLC suffer from various limitations that can be partly overcome by the application of nanomedicines. Nanoparticles is a branch within nanomedicine that can improve the delivery of anticancer drugs, whilst ensuring the stability and sufficient bioavailability following administration. There are many publications available in the literature exploring different types of nanoparticles from different materials. The effectiveness of a treatment option needs to be validated in suitable in vitro and/or in vivo models. This includes the developed nanoparticles, to prove their safety and efficacy. Many researchers have turned towards in vitro models that use normal cells or specific cells from diseased tissues. However, in cellular works, the physiological dynamics that is available in the body could not be mimicked entirely, and hence, there is still possible development of false positive or false negative results from the in vitro models. This article provides an overview of NSCLC, the different nanoparticles available to date, and in vitro evaluation of the nanoparticles. Different types of cells suitable for in vitro study and the important precautions to limit the development of false results are also extensively discussed.
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Affiliation(s)
- Noratiqah Mohtar
- School of Pharmaceutical Sciences, Universiti Sains Malaysia, Gelugor 11800, Penang, Malaysia; (N.M.); (T.P.); (A.M.G.); (C.S.T.); (M.L.T.); (H.A.W.)
| | - Thaigarajan Parumasivam
- School of Pharmaceutical Sciences, Universiti Sains Malaysia, Gelugor 11800, Penang, Malaysia; (N.M.); (T.P.); (A.M.G.); (C.S.T.); (M.L.T.); (H.A.W.)
| | - Amirah Mohd Gazzali
- School of Pharmaceutical Sciences, Universiti Sains Malaysia, Gelugor 11800, Penang, Malaysia; (N.M.); (T.P.); (A.M.G.); (C.S.T.); (M.L.T.); (H.A.W.)
| | - Chu Shan Tan
- School of Pharmaceutical Sciences, Universiti Sains Malaysia, Gelugor 11800, Penang, Malaysia; (N.M.); (T.P.); (A.M.G.); (C.S.T.); (M.L.T.); (H.A.W.)
| | - Mei Lan Tan
- School of Pharmaceutical Sciences, Universiti Sains Malaysia, Gelugor 11800, Penang, Malaysia; (N.M.); (T.P.); (A.M.G.); (C.S.T.); (M.L.T.); (H.A.W.)
| | - Rozana Othman
- Faculty of Pharmacy, Universiti Malaya, Kuala Lumpur 50603, Malaysia
- Center for Natural Products Research and Drug Discovery (CENAR), Universiti Malaya, Kuala Lumpur 50603, Malaysia
| | - Siti Sarah Fazalul Rahiman
- School of Pharmaceutical Sciences, Universiti Sains Malaysia, Gelugor 11800, Penang, Malaysia; (N.M.); (T.P.); (A.M.G.); (C.S.T.); (M.L.T.); (H.A.W.)
| | - Habibah A. Wahab
- School of Pharmaceutical Sciences, Universiti Sains Malaysia, Gelugor 11800, Penang, Malaysia; (N.M.); (T.P.); (A.M.G.); (C.S.T.); (M.L.T.); (H.A.W.)
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Kaberov LI, Kaberova Z, Murmiliuk A, Trousil J, Sedláček O, Konefal R, Zhigunov A, Pavlova E, Vít M, Jirák D, Hoogenboom R, Filippov SK. Fluorine-Containing Block and Gradient Copoly(2-oxazoline)s Based on 2-(3,3,3-Trifluoropropyl)-2-oxazoline: A Quest for the Optimal Self-Assembled Structure for 19F Imaging. Biomacromolecules 2021; 22:2963-2975. [PMID: 34180669 DOI: 10.1021/acs.biomac.1c00367] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
Abstract
The use of fluorinated contrast agents in magnetic resonance imaging (MRI) facilitates improved image quality due to the negligible amount of endogenous fluorine atoms in the body. In this work, we present a comprehensive study of the influence of the amphiphilic polymer structure and composition on its applicability as contrast agents in 19F MRI. Three series of novel fluorine-containing poly(2-oxazoline) copolymers and terpolymers, hydrophilic-fluorophilic, hydrophilic-lipophilic-fluorophilic, and hydrophilic-thermoresponsive-fluorophilic, with block and gradient distributions of the fluorinated units, were synthesized. It was discovered that the CF3 in the 2-(3,3,3-trifluoropropyl)-2-oxazoline (CF3EtOx) group activated the cationic chain end, leading to faster copolymerization kinetics, whereby spontaneous monomer gradients were formed with accelerated incorporation of 2-methyl-2-oxazoline or 2-n-propyl-2-oxazoline with a gradual change to the less-nucleophilic CF3EtOx monomer. The obtained amphiphilic copolymers and terpolymers form spherical or wormlike micelles in water, which was confirmed using transmission electron microscopy (TEM), while small-angle X-ray scattering (SAXS) revealed the core-shell or core-double-shell morphologies of these nanoparticles. The core and shell sizes obey the scaling laws for starlike micelles predicted by the scaling theory. Biocompatibility studies confirm that all copolymers obtained are noncytotoxic and, at the same time, exhibit high sensitivity during in vitro 19F MRI studies. The gradient copolymers provide the best 19F MRI signal-to-noise ratio in comparison with the analogue block copolymer structures, making them most promising as 19F MRI contrast agents.
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Affiliation(s)
- Leonid I Kaberov
- Institute of Macromolecular Chemistry, Czech Academy of Sciences, Heyrovského nám. 2, 162 06 Prague, Czech Republic
| | - Zhansaya Kaberova
- Institute of Macromolecular Chemistry, Czech Academy of Sciences, Heyrovského nám. 2, 162 06 Prague, Czech Republic
| | - Anastasiia Murmiliuk
- Department of Physical and Macromolecular Chemistry, Faculty of Science, Charles University, Hlavova 8, 128 40 Prague, Czech Republic
| | - Jiří Trousil
- Institute of Macromolecular Chemistry, Czech Academy of Sciences, Heyrovského nám. 2, 162 06 Prague, Czech Republic
| | - Ondřej Sedláček
- Department of Physical and Macromolecular Chemistry, Faculty of Science, Charles University, Hlavova 8, 128 40 Prague, Czech Republic.,Supramolecular Chemistry Group, Centre of Macromolecular Chemistry (CMaC), Department of Organic and Macromolecular Chemistry, Ghent University, Krijgslaan 281 S4, B-9000 Ghent, Belgium
| | - Rafal Konefal
- Institute of Macromolecular Chemistry, Czech Academy of Sciences, Heyrovského nám. 2, 162 06 Prague, Czech Republic
| | - Alexander Zhigunov
- Institute of Macromolecular Chemistry, Czech Academy of Sciences, Heyrovského nám. 2, 162 06 Prague, Czech Republic
| | - Ewa Pavlova
- Institute of Macromolecular Chemistry, Czech Academy of Sciences, Heyrovského nám. 2, 162 06 Prague, Czech Republic
| | - Martin Vít
- Faculty of Mechatronics Informatics and Interdisciplinary Studies, Technical University of Liberec, Studentská 1402/2, 461 17 Liberec, Czech Republic
| | - Daniel Jirák
- Institute for Clinical and Experimental Medicine, Vídeňská 9, 140 21 Prague, Czech Republic.,Institute of Biophysics and Informatics, First Faculty of Medicine, Charles University in Prague, Salmovská 1, 120 00 Prague, Czech Republic
| | - Richard Hoogenboom
- Supramolecular Chemistry Group, Centre of Macromolecular Chemistry (CMaC), Department of Organic and Macromolecular Chemistry, Ghent University, Krijgslaan 281 S4, B-9000 Ghent, Belgium
| | - Sergey K Filippov
- Pharmaceutical Sciences Laboratory, Faculty of Science and Engineering, Åbo Akademi University, 20520 Turku, Finland.,Department of Chemistry and Chemical Technology, Al-Farabi Kazakh National University, 050040 Almaty, Kazakhstan
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Yan K, Rawle DJ, Le TT, Suhrbier A. Simple rapid in vitro screening method for SARS-CoV-2 anti-virals that identifies potential cytomorbidity-associated false positives. Virol J 2021; 18:123. [PMID: 34107996 PMCID: PMC8188739 DOI: 10.1186/s12985-021-01587-z] [Citation(s) in RCA: 14] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/10/2021] [Accepted: 05/27/2021] [Indexed: 12/13/2022] Open
Abstract
Background The international SARS-CoV-2 pandemic has resulted in an urgent need to identify new anti-viral drugs for treatment of COVID-19. The initial step to identifying potential candidates usually involves in vitro screening that includes standard cytotoxicity controls. Under-appreciated is that viable, but stressed or otherwise compromised cells, can also have a reduced capacity to replicate virus. A refinement proposed herein for in vitro drug screening thus includes a simple growth assay to identify drug concentrations that cause cellular stress or “cytomorbidity”, as distinct from cytotoxicity or loss of viability. Methods A simple rapid bioassay is presented for antiviral drug screening using Vero E6 cells and inhibition of SARS-CoV-2 induced cytopathic effects (CPE) measured using crystal violet staining. We use high cell density for cytotoxicity assays, and low cell density for cytomorbidity assays. Results The assay clearly illustrated the anti-viral activity of remdesivir, a drug known to inhibit SARS-CoV-2 replication. In contrast, nitazoxanide, oleuropein, cyclosporine A and ribavirin all showed no ability to inhibit SARS-CoV-2 CPE. Hydroxychloroquine, cyclohexamide, didemnin B, γ-mangostin and linoleic acid were all able to inhibit viral CPE at concentrations that did not induce cytotoxicity. However, these drugs inhibited CPE at concentrations that induced cytomorbidity, indicating non-specific anti-viral activity. Conclusions We describe the methodology for a simple in vitro drug screening assay that identifies potential anti-viral drugs via their ability to inhibit SARS-CoV-2-induced CPE. The additional growth assay illustrated how several drugs display anti-viral activity at concentrations that induce cytomorbidity. For instance, hydroxychloroquine showed anti-viral activity at concentrations that slow cell growth, arguing that its purported in vitro anti-viral activity arises from non-specific impairment of cellular activities. The cytomorbidity assay can therefore rapidly exclude potential false positives. Supplementary Information The online version contains supplementary material available at 10.1186/s12985-021-01587-z.
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Affiliation(s)
- Kexin Yan
- QIMR Berghofer Medical Research Institute, Brisbane, QLD, 4029, Australia
| | - Daniel J Rawle
- QIMR Berghofer Medical Research Institute, Brisbane, QLD, 4029, Australia
| | - Thuy T Le
- QIMR Berghofer Medical Research Institute, Brisbane, QLD, 4029, Australia
| | - Andreas Suhrbier
- QIMR Berghofer Medical Research Institute, Brisbane, QLD, 4029, Australia. .,Australian Infectious Disease Research Centre, GVN Center of Excellence, Brisbane, QLD, 4029 and 4072, Australia.
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Ihara M, Zhang H, Ihara MO, Kato D, Tanaka H. Proposal for fluorescence-based in vitro assay using human and zebrafish monoamine transporters to detect biological activities of antidepressants in wastewater. THE SCIENCE OF THE TOTAL ENVIRONMENT 2021; 770:144665. [PMID: 33513512 DOI: 10.1016/j.scitotenv.2020.144665] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 09/24/2020] [Revised: 12/11/2020] [Accepted: 12/17/2020] [Indexed: 06/12/2023]
Abstract
Antidepressants are among the most commonly detected pharmaceuticals in the aquatic environment. As they modulate neurotransmission in nervous systems, behavioural abnormalities among aquatic species are of concern. It is possible to measure the concentrations of selected antidepressants by chemical analysis, but other non-target antidepressants and active metabolites might also be present. Here, we propose an "in vitro monoamine transporter inhibition assay" to measure the biological activity of antidepressants, particularly monoamine transporter inhibitors, in wastewater. We used APP, a fluorescent substrate for monoamine transporters, to measure the activity of wastewater extracts at inhibiting APP uptake through the human serotonin transporter (hSERT), norepinephrine transporter (hNET), and dopamine transporter, and the zebrafish SERT (zSERT). We confirmed that the assay could measure the biological activity of test antidepressants. Interestingly, the IC50 values of antidepressants (the concentration that gave a 50% reduction of APP uptake) for the zSERT were smaller than those for the hSERT. For example, IC50 value of desipramine for the zSERT was 1/200 of that for the hSERT. These results indicate that antidepressants inhibited zSERT more strongly than hSERT. Then we applied the assay to extracts of effluent from municipal wastewater treatment plants and detected biological activity of antidepressants specifically against the hSERT, hNET, and zSERT for the first time. For the hSERT, antidepressant-equivalent quantities (EQs) ranged from 2.2 × 101 to 2.5 × 102 ng-clomipramine-EQ/L. For the hNET, EQs ranged from below limit of detection to 8.2 × 101 ng-desipramine-EQ/L. For the zSERT, EQs ranged from 2.8 × 102 to 3.3 × 102 ng-duloxetine-EQ/L. The in vitro monoamine transporter inhibition assay is thus useful for measuring the biological activity of antidepressants in the aquatic environment.
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Affiliation(s)
- Masaru Ihara
- Research Center for Environmental Quality Management, Kyoto University, 1-2 Yumihama, Otsu, Shiga 520-0811, Japan.
| | - Han Zhang
- Research Center for Environmental Quality Management, Kyoto University, 1-2 Yumihama, Otsu, Shiga 520-0811, Japan
| | - Mariko O Ihara
- Research Center for Environmental Quality Management, Kyoto University, 1-2 Yumihama, Otsu, Shiga 520-0811, Japan
| | - Daisuke Kato
- Research Center for Environmental Quality Management, Kyoto University, 1-2 Yumihama, Otsu, Shiga 520-0811, Japan
| | - Hiroaki Tanaka
- Research Center for Environmental Quality Management, Kyoto University, 1-2 Yumihama, Otsu, Shiga 520-0811, Japan
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Nataraj BH, Ramesh C, Mallappa RH. Extractable surface proteins of indigenous probiotic strains confer anti-adhesion knack and protect against methicillin-resistant Staphylococcus aureus induced epithelial hyperpermeability in HT-29 cell line. Microb Pathog 2021; 158:104974. [PMID: 34015494 DOI: 10.1016/j.micpath.2021.104974] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/14/2021] [Revised: 04/29/2021] [Accepted: 04/30/2021] [Indexed: 11/28/2022]
Abstract
Probiotic intervention has been long believed to have beneficial effects on human health by curbing the intestinal colonization of pathogens. However, the application of live probiotics therapy may not be an ideal approach to circumvent the infections of superbug origin due to the risk of horizontal antibiotic resistance genes transfer. In this study, the anti-adhesion ability of extractable cell surface proteins from two indigenous potential probiotic strains (Lactiplantibacillus plantarum A5 and Limosilactobacillus fermentum Lf1) and two standard reference strains (Lactobacillus acidophilus NCFM and Lacticaseibacillus rhamnosus LGG) was evaluated against clinical isolates of Methicillin-Resistant Staphylococcus aureus (MRSA) on porcine gastric mucin and HT-29 cells. The surface proteins from the probiotic strains were extracted by treatment with 5 M lithium chloride. The surface protein quantification and SDS-PAGE profiling indicated that the yield and protein patterns were strain-specific. Surface proteins significantly hampered the mucoadhesion of MRSA isolates via protective, competitive, and displacement. Similarly, the treatment with surface proteins probiotic strains displayed anti-adhesion against MRSA isolates on HT-29 cells without affecting the viability of the cell line. Surface proteins treatment to the confluent monolayer of HT-29 cells maintained the epithelial integrity; however, MRSA isolates (109 cells/mL) showed considerable alteration in the epithelial integrity by exacerbating the FITC-dextran transflux. Contrarily, the co-treatment with surface proteins with MRSA isolates significantly lowered the FITC-dextran transflux across the differentiated HT-29 monolayer. Overall, the findings of this study suggest that probiotic-derived surface proteins could be the novel biotherapeutics to combat the MRSA colonization and their concomitant intestinal infections.
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Affiliation(s)
| | - Chette Ramesh
- Molecular Biology Unit, Dairy Microbiology Division, ICAR-National Dairy Research Institute, Karnal, 132001, Haryana, India
| | - Rashmi Hogarehalli Mallappa
- Molecular Biology Unit, Dairy Microbiology Division, ICAR-National Dairy Research Institute, Karnal, 132001, Haryana, India.
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Wu CP, Hung TH, Lusvarghi S, Chu YH, Hsiao SH, Huang YH, Chang YT, Ambudkar SV. The third-generation EGFR inhibitor almonertinib (HS-10296) resensitizes ABCB1-overexpressing multidrug-resistant cancer cells to chemotherapeutic drugs. Biochem Pharmacol 2021; 188:114516. [PMID: 33713643 DOI: 10.1016/j.bcp.2021.114516] [Citation(s) in RCA: 26] [Impact Index Per Article: 6.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/22/2020] [Revised: 03/03/2021] [Accepted: 03/04/2021] [Indexed: 02/07/2023]
Abstract
The overexpression of the human ATP-binding cassette (ABC) drug transporter ABCB1 (P-glycoprotein, P-gp) or ABCG2 (breast cancer resistance protein, BCRP) in cancer cells often contributes significantly to the development of multidrug resistance (MDR) in cancer patients. Previous reports have demonstrated that some epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) could modulate the activity of ABCB1 and/or ABCG2 in human cancer cells, whereas some EGFR TKIs are transport substrates of these transporters. Almonertinib (HS-10296) is a promising, orally available third-generation EGFR TKI for the treatment of EGFR T790M mutation-positive non-small cell lung cancer (NSCLC) in patients who have progressed on or after other EGFR TKI therapies. Additional clinical trials are currently in progress to study almonertinib as monotherapy and in combination with other agents in patients with NSCLC. In the present work, we found that neither ABCB1 nor ABCG2 confers significant resistance to almonertinib. More importantly, we discovered that almonertinib was able to reverse MDR mediated by ABCB1, but not ABCG2, in multidrug-resistant cancer cells at submicromolar concentrations by inhibiting the drug transport activity of ABCB1 without affecting its expression level. These findings are further supported by in silico docking of almonertinib in the drug-binding pocket of ABCB1. In summary, our study revealed an additional activity of almonertinib to re-sensitize ABCB1-overexpressing multidrug-resistant cancer cells to conventional chemotherapeutic drugs, which may be beneficial for cancer patients and warrant further investigation.
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Affiliation(s)
- Chung-Pu Wu
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan; Department of Physiology and Pharmacology, and College of Medicine, Chang Gung University, Tao-Yuan, Taiwan; Department of Obstetrics and Gynecology, Taipei Chang Gung Memorial Hospital, Taipei, Taiwan.
| | - Tai-Ho Hung
- Department of Chinese Medicine, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan; Department of Obstetrics and Gynecology, Taipei Chang Gung Memorial Hospital, Taipei, Taiwan
| | - Sabrina Lusvarghi
- Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD, United States
| | - Yi-Hsuan Chu
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan
| | - Sung-Han Hsiao
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan
| | - Yang-Hui Huang
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan; Department of Physiology and Pharmacology, and College of Medicine, Chang Gung University, Tao-Yuan, Taiwan
| | - Yu-Tzu Chang
- Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan; Department of Physiology and Pharmacology, and College of Medicine, Chang Gung University, Tao-Yuan, Taiwan
| | - Suresh V Ambudkar
- Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, MD, United States
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