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Tomsuk Ö, Kaçar S. Mechanistic Insights into Silymarin-Induced Apoptosis and Growth Inhibition in SPC212 Human Mesothelioma Cells. Cell Biochem Biophys 2025; 83:2405-2414. [PMID: 39747779 DOI: 10.1007/s12013-024-01650-w] [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] [Accepted: 12/17/2024] [Indexed: 01/04/2025]
Abstract
Silymarin, a flavonoid complex isolated from Silybum marianum, possesses various biological properties, including antioxidant, anti-inflammatory, anti-glycation, and hepatoprotective effects. In the present study, we investigated the effects of silymarin on the SPC212 human mesothelioma cell line. MTT and neutral red assays were performed to examine the cytotoxic effects of silymarin. The apoptotic effect was investigated using AO/EB and DAPI staining, and morphological changes were observed using H&E and May-Grünwald staining. Additionally, immunocytochemistry was performed to detect Bax, Bcl2, and PCNA. Our results indicated that silymarin has a dose-dependent cytotoxic effect on SPC212 cells, with an IC50 value of approximately 187.5 µM. Silymarin induces apoptotic hallmarks such as apoptotic bodies, cell shrinkage, and nuclear condensation. In conclusion, silymarin demonstrated cytotoxic and apoptotic effects as well as morphological changes in SPC212 human mesothelioma cells. Further detailed studies are warranted to explore the potential of silymarin as an anti-cancer agent.
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Affiliation(s)
- Özlem Tomsuk
- Graduate School of Natural and Applied Sciences, Biotechnology and Biosafety Department, Eskişehir Osmangazi University, Eskişehir, Türkiye.
- Cellular Therapy and Stem Cell Production Application and Research Center (ESTEM), Eskişehir Osmangazi University, Eskişehir, Türkiye.
- Faculty of Medicine, Department of Histology and Embryology, Eskişehir Osmangazi University, Eskişehir, Türkiye.
| | - Sedat Kaçar
- Faculty of Medicine, Department of Histology and Embryology, Eskişehir Osmangazi University, Eskişehir, Türkiye
- Vera Bradley Foundation Center for Breast Cancer Research, Department of Surgery & Division of Oncologic Surgery, Indiana University School of Medicine, Indianapolis, USA
- Division of Pulmonary, Critical Care, Sleep and Occupational Medicine, Department of Medicine, Indiana University, Indianapolis, Indiana, USA
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El-Moslamy SH, El-Maradny YA, El-Sayed MH, El-Sakhawy MA, El-Fakharany EM. Facile phyto-mediated synthesis of ternary CuO/Mn 3O 4/ZnO nanocomposite using Nigella Sativa seeds extract: characterization,antimicrobial, and biomedical evaluations. Sci Rep 2025; 15:16139. [PMID: 40341630 PMCID: PMC12062449 DOI: 10.1038/s41598-024-85044-1] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/21/2024] [Accepted: 12/30/2024] [Indexed: 05/10/2025] Open
Abstract
The phyto-synthesis of ternary CuO/ Mn3O4/ZnO nanocomposite was achieved by the utilization of an eco-friendly, straightforward approach that involved the extract of Nigella sativa seeds. Our ternary nanocomposite appears to include equal amounts of CuO, Mn3O4, and ZnO based on the atomic percentages. The results indicate that a robust and thermally stable CuO/Mn3O4/ZnO nanocomposite was developed in stable nanosuspensions. The CuO/Mn3O4/ZnO nanocomposites showed antimicrobial capabilities against multidrug-resistant human pathogens. The highest biofilm reduction in viable planktonic populations of all human pathogens investigated was significantly reduced by the CuO/Mn3O4/ZnO ternary nanocomposites with a value of 18.5 µg/mL. The unique, enhanced, and triple-combined properties enabled the nanocomposite to have strong antimicrobial ability. The CuO/Mn3O4/ZnO nanocomposite exhibited strong anticancer activity against A549, MDA, HCT-116, and HepG2 cells, with selectivity index values ranging from 24.72 to 41.96. The CuO/Mn3O4/ZnO nanocomposite appeared to induce selective dose-dependent nuclear condensation and cell shrinkage in the treated cancer cells, significantly inducing the apoptosis mechanism to combat cancer progression. The phytosynthetic CuO/Mn3O4/ZnO nanocomposite appears to induce selective dose-dependent nuclear condensation and cell shrinkage in treated cancer cells, significantly triggering apoptotic mechanisms to combat cancer progression. This apoptotic pathway was confirmed by the strong affinity of CuO/Mn3O4/ZnO nanocomposites for ErbBs and VEGF with potent antioxidant activity to scavenge ABTS and DPPH radicals at EC50 values of 236.6 µg/mL and 134.8 µg/mL, respectively.
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Affiliation(s)
- Shahira H El-Moslamy
- Department of Bioprocess Development, Genetic Engineering and Biotechnology Research Institute (GEBRI), City of Scientific Research and Technological Applications (SRTA-city), New Borg El Arab City, 21934, Alexandria, Egypt.
| | - Yousra A El-Maradny
- Pharmaceutical and Fermentation Industries Development Centre (PFIDC), City of Scientific Research and Technological Applications (SRTA-City), New Borg Al-Arab City, 21934, Alexandria, Egypt
| | - Mohamed H El-Sayed
- Department of Biology, College of Sciences and Arts-Rafha, Northern Border University, Arar, Saudi Arabia
| | - Mohamed A El-Sakhawy
- Department of Medical Laboratory, College of Applied Medical Sciences, Prince Sattam bin Abdulaziz University, Al-Kharj, 11942, Saudi Arabia
- Department of Medicinal and Aromatic Plants, Desert Research Center, Cairo, Egypt
| | - Esmail M El-Fakharany
- Pharmaceutical and Fermentation Industries Development Centre (PFIDC), City of Scientific Research and Technological Applications (SRTA-City), New Borg Al-Arab City, 21934, Alexandria, Egypt.
- Protein Research Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), City of Scientific Research and Technological Applications (SRTA-City), New Borg El-Arab City, 21934, Alexandria, Egypt.
- Pharos University in Alexandria, Canal El Mahmoudia Street, Beside Green Plaza Complex, 21648, Alexandria, Egypt.
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Al-Haideri M. Silymarin suppresses proliferation and PD-L1 expression in colorectal cancer cells and increases inflammatory CD8+ cells in tumor-bearing mice. Clin Res Hepatol Gastroenterol 2024; 48:102425. [PMID: 39048076 DOI: 10.1016/j.clinre.2024.102425] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/13/2024] [Revised: 06/15/2024] [Accepted: 07/20/2024] [Indexed: 07/27/2024]
Abstract
INTRODUCTION Silymarin as an herbal medicine has shown anticancer effects on tumor cells, while having low toxicity in normal cells. In this study, the effects of Silymarin on proliferation and apoptosis of colorectal cancer cells and its impact on immune response against cancer cells were evaluated in vitro and in vivo. METHODS AND MATERIALS The effect of Silymarin on CT-26 and Caco-2 cells proliferation and apoptosis were demonstrated by MTT assay and PI staining. A subcutaneous tumor of colorectal cancer was developed. Silymarin and Doxorubicin were administrated by intravenous injection. qRT-PCR analyses was performed on blood samples and tumor tissues. Spleen tissue was used to evaluate CD8+ T cell immune responses. Histological study was carried out on tumor tissues. RESULTS Silymarin showed anti-proliferative effects on CT-26 and Caco-2 cells. The markers of immunogenic cell death (Calreticulin exposure, ATP secretion, and HMGB1 secretion) significantly increased in both cell lines in the presence of silymarin. The expression of genes related to cell proliferation particularly β-Catenin and Cycline D1, and also anti-apoptotic ones such as Bcl-2 significantly reduced in mice treated with Silymarin while the expression of pro-apoptotic Bax increased. The RNA level of PD-L1 decreased in tumor tissues exposed by Silymarin. Moreover, the number of CTLs increased in the spleen of mice treated with Silymarin in comparison with untreated mice. Decreased tumor size and also survival of colorectal cancer cells in Silymarin-treated mice were observed in histological analysis. CONCLUSION Silymarin treatment showed a suppressive role on colorectal cancer cells almost as much as Doxorubicin. Our study indicated that having a low toxicity profile, cost-effectiveness, and availability of raw materials, plant-derived Silymarin can be a good candidate for further investigation to treat CRC.
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Affiliation(s)
- Maysoon Al-Haideri
- School of medicine, Pharmacy Department, University of Kurdistan Hawlêr, Erbil, Kurdistan, Iraq.
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Lee B, Kim YY, Jeong S, Lee SW, Lee SJ, Rho MC, Kim SH, Lee S. Oleanolic Acid Acetate Alleviates Cisplatin-Induced Nephrotoxicity via Inhibition of Apoptosis and Necroptosis In Vitro and In Vivo. TOXICS 2024; 12:301. [PMID: 38668524 PMCID: PMC11054587 DOI: 10.3390/toxics12040301] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 03/14/2024] [Revised: 04/12/2024] [Accepted: 04/16/2024] [Indexed: 04/29/2024]
Abstract
Cisplatin is a widely used anti-cancer drug for treating solid tumors, but it is associated with severe side effects, including nephrotoxicity. Various studies have suggested that the nephrotoxicity of cisplatin could be overcome; nonetheless, an effective adjuvant drug has not yet been established. Oleanolic acid acetate (OAA), a triterpenoid isolated from Vigna angularis, is commonly used to treat inflammatory and allergic diseases. This study aimed to investigate the protective effects of OAA against cisplatin-induced apoptosis and necroptosis using TCMK-1 cells and a mouse model. In cisplatin-treated TCMK-1 cells, OAA treatment significantly reduced Bax and cleaved-caspase3 expression, whereas it increased Bcl-2 expression. Moreover, in a cisplatin-induced kidney injury mouse model, OAA treatment alleviated weight loss in the body and major organs and also relieved cisplatin-induced nephrotoxicity symptoms. RNA sequencing analysis of kidney tissues identified lipocalin-2 as the most upregulated gene by cisplatin. Additionally, necroptosis-related genes such as receptor-interacting protein kinase (RIPK) and mixed lineage kinase domain-like (MLKL) were identified. In an in vitro study, the phosphorylation of RIPKs and MLKL was reduced by OAA pretreatment in both cisplatin-treated cells and cells boosted via co-treatment with z-VAD-FMK. In conclusion, OAA could protect the kidney from cisplatin-induced nephrotoxicity and may serve as an anti-cancer adjuvant.
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Affiliation(s)
- Bori Lee
- Functional Biomaterials Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56212, Republic of Korea; (B.L.); (Y.-Y.K.); (S.J.); (S.W.L.); (S.-J.L.); (M.-C.R.)
| | - Yeon-Yong Kim
- Functional Biomaterials Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56212, Republic of Korea; (B.L.); (Y.-Y.K.); (S.J.); (S.W.L.); (S.-J.L.); (M.-C.R.)
| | - Seungwon Jeong
- Functional Biomaterials Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56212, Republic of Korea; (B.L.); (Y.-Y.K.); (S.J.); (S.W.L.); (S.-J.L.); (M.-C.R.)
| | - Seung Woong Lee
- Functional Biomaterials Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56212, Republic of Korea; (B.L.); (Y.-Y.K.); (S.J.); (S.W.L.); (S.-J.L.); (M.-C.R.)
| | - Seung-Jae Lee
- Functional Biomaterials Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56212, Republic of Korea; (B.L.); (Y.-Y.K.); (S.J.); (S.W.L.); (S.-J.L.); (M.-C.R.)
| | - Mun-Chual Rho
- Functional Biomaterials Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56212, Republic of Korea; (B.L.); (Y.-Y.K.); (S.J.); (S.W.L.); (S.-J.L.); (M.-C.R.)
| | - Sang-Hyun Kim
- Cell and Matrix Research Institute, Department of Pharmacology, School of Medicine, Kyungpook National University, Daegu 41944, Republic of Korea
| | - Soyoung Lee
- Functional Biomaterials Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup 56212, Republic of Korea; (B.L.); (Y.-Y.K.); (S.J.); (S.W.L.); (S.-J.L.); (M.-C.R.)
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Wang Y, Yuan AJ, Wu YJ, Wu LM, Zhang L. Silymarin in cancer therapy: Mechanisms of action, protective roles in chemotherapy-induced toxicity, and nanoformulations. J Funct Foods 2023. [DOI: 10.1016/j.jff.2022.105384] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/24/2022] Open
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Non-Canonical Programmed Cell Death in Colon Cancer. Cancers (Basel) 2022; 14:cancers14143309. [PMID: 35884370 PMCID: PMC9320762 DOI: 10.3390/cancers14143309] [Citation(s) in RCA: 13] [Impact Index Per Article: 4.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/13/2022] [Revised: 06/24/2022] [Accepted: 07/05/2022] [Indexed: 12/18/2022] Open
Abstract
Simple Summary Non-canonical PCD is an important player in colon cancer cell suicide. It influences colon cancer in many ways, such as through tumorigenesis, treatment, and prognosis. In this review, we present the mechanism, application, and prospect of different types of non-canonical PCD in colon cancer. Abstract Programmed cell death (PCD) is an evolutionarily conserved process of cell suicide that is regulated by various genes and the interaction of multiple signal pathways. Non-canonical programmed cell death (PCD) represents different signaling excluding apoptosis. Colon cancer is the third most incident and the fourth most mortal worldwide. Multiple factors such as alcohol, obesity, and genetic and epigenetic alternations contribute to the carcinogenesis of colon cancer. In recent years, emerging evidence has suggested that diverse types of non-canonical programmed cell death are involved in the initiation and development of colon cancer, including mitotic catastrophe, ferroptosis, pyroptosis, necroptosis, parthanatos, oxeiptosis, NETosis, PANoptosis, and entosis. In this review, we summarized the association of different types of non-canonical PCD with tumorigenesis, progression, prevention, treatments, and prognosis of colon cancer. In addition, the prospect of drug-resistant colon cancer therapy related to non-canonical PCD, and the interaction between different types of non-canonical PCD, was systemically reviewed.
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Mi XJ, Choi HS, Perumalsamy H, Shanmugam R, Thangavelu L, Balusamy SR, Kim YJ. Biosynthesis and cytotoxic effect of silymarin-functionalized selenium nanoparticles induced autophagy mediated cellular apoptosis via downregulation of PI3K/Akt/mTOR pathway in gastric cancer. PHYTOMEDICINE : INTERNATIONAL JOURNAL OF PHYTOTHERAPY AND PHYTOPHARMACOLOGY 2022; 99:154014. [PMID: 35247670 DOI: 10.1016/j.phymed.2022.154014] [Citation(s) in RCA: 32] [Impact Index Per Article: 10.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 08/10/2021] [Revised: 02/08/2022] [Accepted: 02/25/2022] [Indexed: 06/14/2023]
Abstract
BACKGROUND Silymarin, a blend of flavonolignans isolated from plant Silybum marianum L., has long been used as an herbal medicine. Biogenic routes especially the plant-based synthesis of selenium nanoparticles (SeNPs) is safe, eco-friendly, nontoxic and being considered as one of the best strategies for treatment of cancer. PURPOSE Silymarin-mediated green synthesis of SeNPs and their possibility as an anticancer agent have not been reported to date. Therefore, our present study was aimed to synthesize and characterize the selenium mediated silymarin nanoparticles (Si-SeNPs) from silymarin and investigate their possibility as an anticancer agent. METHODS The physicochemical characteristics of Si-SeNPs were analyzed using various analytical techniques, such as HPLC, field emission-transmission electron microscope, energy-dispersive X-ray spectrometer, and thermogravimetric analysis. The underlying molecular mechanism were evaluated using AGS gastric cancer cells. RESULTS Compared with silymarin, the Si-SeNPs exhibited significantly increased cytotoxic effect of AGS cells without exhibiting toxicity on normal cells. Real time PCR and western blotting analysis indicated that Si-SeNPs induced expression of Bax/Bcl-2, cytochrome c, and cleavage of caspase proteins, which is associated with mitochondria-mediated apoptosis signaling in AGS cells. Moreover, agonist assay using PI3K activator indicated that Si-SeNPs-inhibited PI3K/AKT/mTOR pathways were significantly associated as an autophagy and apoptosis signaling in AGS cells. CONCLUSION Our study demonstrated the improved anticancer efficacy of Si-SeNPs- induced apoptosis and autophagy pathways, and therefore recommended Si-SeNPs as a novel anticancer agent after in vivo studies.
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Affiliation(s)
- Xiao-Jie Mi
- Graduate School of Biotechnology, and College of Life Science, Kyung Hee University, Yongin-si, 17104, Gyeonggi-do, Republic of Korea
| | - Han Sol Choi
- Graduate School of Biotechnology, and College of Life Science, Kyung Hee University, Yongin-si, 17104, Gyeonggi-do, Republic of Korea
| | - Haribalan Perumalsamy
- Graduate School of Biotechnology, and College of Life Science, Kyung Hee University, Yongin-si, 17104, Gyeonggi-do, Republic of Korea; Research Institute for Convergence of Basic Science, Hanyang University, Seoul 04763, Republic of Korea
| | - Rajeshkumar Shanmugam
- Department of Pharmacology, Saveetha Dental College and Hospitals, Saveetha University, SIMATS, Chennai 600077, TN, India
| | - Lakshmi Thangavelu
- Department of Pharmacology, Saveetha Dental College and Hospitals, Saveetha University, SIMATS, Chennai 600077, TN, India
| | - Sri Renukadevi Balusamy
- Department of Food Science and Biotechnology, Sejong University, Gwangjin-gu, Seoul, 05006, Republic of Korea.
| | - Yeon-Ju Kim
- Graduate School of Biotechnology, and College of Life Science, Kyung Hee University, Yongin-si, 17104, Gyeonggi-do, Republic of Korea.
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Valizade M, Raesi Vanani A, Rezaei M, Khorsandi LS, Zeidooni L, Mahdavinia M. Mesobuthus eupeus venom induced injury in the colorectal carcinoma cell line (HT29) through altering the mitochondria membrane stability. IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES 2020; 23:760-767. [PMID: 32695292 PMCID: PMC7351445 DOI: 10.22038/ijbms.2020.40884.9659] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Indexed: 11/09/2022]
Abstract
Objective(s): The purpose of this study was to investigate cytotoxicity and membrane toxicity effects induced by Mesobuthus eupeus venom (MEV) on the HT-29 cell line. Materials and Methods: To determine the in vitro cytotoxicity via MTT assays, HT-29 (as cancer cell line) and Hek-293T (as normal cell) were treated through different concentrations of MEV, and cytotoxicity effects were then measured through assessment of mitochondrial membrane potential (ΔΨm), reactive oxygen species (ROS) generation, and apoptosis induction. The colony formation assay was performed to measure the antiproliferative effect of MEV on HT-29 cells. Nuclei alterations were also observed during apoptosis following DAPI staining. Besides, atomic force microscopy (AFM) was used to detect alterations in morphology and ultrastructure of the cells at a nanoscale level. Results: According to MTT and clonogenic assays, MEV caused a significant decrease in cell viability and proliferation of HT-29 cells while it did not have any impact on normal cells and the IC50 value was found to be 10 µg/ml. Induction of apoptosis was also confirmed by flowcytometric analysis in HT-29 cells. Moreover, the results indicated that MEV had led to a suppression of proliferation and induction of apoptosis through increased ROS and depolarization of mitochondria. Furthermore, AFM imaging demonstrated apoptosis cell death after being treated with MEV in HT-29 cells. Conclusion: This study showed that MEV had an antiproliferative effect on HT-29 cells by inducing apoptosis through the mitochondria signaling pathway. These findings suggested that MEV could be used as a promising natural remedy for cancer treatment.
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Affiliation(s)
- Massood Valizade
- Cell & Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.,Toxicology Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
| | - Atefeh Raesi Vanani
- Toxicology Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.,Department of Toxicology, School of Pharmacy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
| | - Mohsen Rezaei
- Toxicology Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.,Department of Toxicology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran
| | - Laya Sadat Khorsandi
- Cell & Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.,Department of Anatomical Sciences, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
| | - Leila Zeidooni
- Toxicology Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
| | - Masoud Mahdavinia
- Toxicology Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.,Department of Toxicology, School of Pharmacy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran
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Qu D, Ma J, Song N, Hui L, Yang L, Guo Y, Sang C. Lappaconitine sulfate induces apoptosis and G0/G1 phase cell cycle arrest by PI3K/AKT signaling pathway in human non-small cell lung cancer A549 cells. Acta Histochem 2020; 122:151557. [PMID: 32622431 DOI: 10.1016/j.acthis.2020.151557] [Citation(s) in RCA: 12] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/26/2020] [Revised: 04/27/2020] [Accepted: 05/02/2020] [Indexed: 02/09/2023]
Abstract
Lappaconitine sulfate (LS) has good solubility and bioavailability. We have previously studied the anti-proliferative activity of LS on colon cancer HT-29 cell, but its anti-proliferative activity and molecular mechanism on human non-small cell lung cancer A549 cells are still unclear. This study was to investigate the effects of LS on proliferation, cell cycle and apoptosis in human non-small cell lung cancer A549 cells, and its possible molecular mechanisms. Cell proliferation activity was measured by Cell Counting Kit-8 (CCK-8) and 5-Ethynyl-2'- deoxyuridine (EdU) cell proliferation kit. Cell cycle was detected by propidium iodide (PI) flow cytometry. Apoptosis was detected by Annexin-V-FITC/PI method. Western blot was used to detect cycle and apoptosis-related proteins expression. These results showed that the proliferation activity of LS was significantly decreased in A549 cells, showing a dose- and time-dependent manner (p < 0.05). LS could increase the proportion of G0/G1 phase cells and decrease the proportion of cells in S phase, showing obvious G0/G1 phase arrest. LS significantly inhibited the expression of p-PI3K/PI3K, p-AKT/AKT, Cyclin D1 and Bcl-2 proteins (p < 0.05), and increased the expression of p53, p21, Bax, caspase 3 and caspase 9 (p < 0.05). Moreover, PI3K inhibitor (LY294002) significantly decreased A549 cell viability rate induced by LS, abrogated the activation of p-PI3K/PI3K and p-AKT/AKT in the presence of LS. These results indicated that LS could block A549 cells in the G0/G1 phase, induce apoptosis, and inhibit cell proliferation through the PI3K/AKT signaling pathway.
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Maruszewska A, Tarasiuk J. Quercetin Triggers Induction of Apoptotic and Lysosomal Death of Sensitive and Multidrug Resistant Leukaemia HL60 Cells. Nutr Cancer 2020; 73:484-501. [PMID: 32329631 DOI: 10.1080/01635581.2020.1752745] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/11/2023]
Abstract
Multidrug resistance (MDR) constitutes the major cause of the failure in anticancer therapy. One of the most important mechanisms leading to the occurrence of MDR is related to the modulation of cellular death pathways. The aim of this study was to determine the effect of quercetin (Q) on triggering the programed death of human promyelocytic leukemia sensitive cells HL60 as well as multidrug resistant HL60/VINC cells overexpressing P-glycoprotein and HL60/MX2 cells characterized by the presence of mutated α isoform of topoisomerase II and the absence of β isoform of this enzyme. Q exerted comparable cytotoxic activities toward sensitive HL60 cells and their MDR counterparts. It was also found that this compound modulated the cellular level of reactive oxygen species (ROS) and led to the marked decrease in cellular GSH level. Furthermore, it was demonstrated that Q used at IC50 and IC90 significantly increased the percentage of sub-G1 subpopulation of all studied leukemia cells causing oligonucleosomal DNA fragmentation. The present study also indicated that Q used at IC90 triggers predominantly programed cell death of sensitive HL60 cells and their MDR counterparts by induction of apoptosis occurring with the involvement of caspase-3 and caspase-8 as well as by lysosome membrane permeabilization-dependent mechanisms.
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Affiliation(s)
- Agnieszka Maruszewska
- Department of Biochemistry, Faculty of Biology, University of Szczecin, Szczecin, Poland.,Molecular Biology and Biotechnology Center, Faculty of Biology, University of Szczecin, Szczecin, Poland
| | - Jolanta Tarasiuk
- Department of Biochemistry, Faculty of Biology, University of Szczecin, Szczecin, Poland.,Molecular Biology and Biotechnology Center, Faculty of Biology, University of Szczecin, Szczecin, Poland
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Girardi B, Pricci M, Giorgio F, Piazzolla M, Iannone A, Losurdo G, Principi M, Barone M, Ierardi E, Di Leo A. Silymarin, boswellic acid and curcumin enriched dietetic formulation reduces the growth of inherited intestinal polyps in an animal model. World J Gastroenterol 2020; 26:1601-1612. [PMID: 32327909 PMCID: PMC7167411 DOI: 10.3748/wjg.v26.i14.1601] [Citation(s) in RCA: 12] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 12/07/2019] [Revised: 03/06/2020] [Accepted: 03/27/2020] [Indexed: 02/06/2023] Open
Abstract
BACKGROUND Some substances of plant origin have been reported to exert an effect in reducing intestinal neoplasm development, especially in animal models. Adenomatous polyposis coli multiple intestinal neoplasia - ApcMin/+ is the most studied murine model of genetic intestinal carcinogenesis. AIM To assess whether an enriched nutritional formulation (silymarin, boswellic acid and curcumin) with proven "in vitro" and "in vivo" anti-carcinogenetic properties may prevent inherited intestinal cancer in animal model. METHODS Forty adenomatous polyposis coli multiple intestinal neoplasia - ApcMin/+ mice were used for the study of cancer prevention. They were divided into two groups: 20 assumed standard and 20 enriched diet. At the 110th d animals were sacrificed. In each group, four subgroups received intraperitoneal bromodeoxyuridine injection at different times (24, 48, 72 and 96 h before the sacrifice) in order to assess epithelial turnover. Moreover, we evaluated the following parameters: Intestinal polypoid lesion number and size on autoptic tissue, dysplasia and neoplasia areas by histological examination of the whole small intestine, inflammation by histology and cytokine mRNA expression by real-time polymerase chain reaction, bromodeoxyuridine and TUNEL immuno-fluorescence for epithelial turnover and apoptosis, respectively. Additionally, we performed western blotting analysis for the expression of estrogen alpha and beta receptors, cyclin D1 and cleaved caspase 3 in normal and polypoid tissues. RESULTS Compared to standard, enriched diet reduced the total number (203 vs 416) and the mean ± SD/animal (12.6 ± 5.0 vs 26.0 ± 8.8; P < 0.001) of polypoid lesions. In enriched diet group a reduction in polyp size was observed (P < 0.001). Histological inflammation and pro-inflammatory cytokine expression were similar in both groups. Areas of low-grade dysplasia (P < 0.001) and intestinal carcinoma (IC; P < 0.001) were significantly decreased in enriched diet group. IC was observed in 100% in standard and 85% in enriched formulation assuming animals. Enriched diet showed a faster epithelial migration and an increased apoptosis in normal mucosa and low-grade dysplasia areas (P < 0.001). At western blotting, estrogen receptor beta protein was well expressed in normal mucosa of enriched and standard groups, with a more marked trend associated to the first one. Estrogen receptor alpha was similarly expressed in normal and polypoid mucosa of standard and enriched diet group. Cleaved caspase 3 showed in normal mucosa a stronger signal in enriched than in standard diet. Cyclin D1 was more expressed in standard than enriched diet group of both normal and polypoid tissue. CONCLUSION Our results are suggestive of a chemo-preventive synergic effect of the components (silymarin, boswellic acid and curcumin) of an enriched formulation in inherited IC. This effect may be mediated by the reduction of epithelial proliferation, the increase of apoptosis and the acceleration of villous cell renewal due to dietary formulation intake.
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Affiliation(s)
| | | | | | - Mariano Piazzolla
- Gastroenterology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari 70124, Italy
| | - Andrea Iannone
- Gastroenterology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari 70124, Italy
| | - Giuseppe Losurdo
- Gastroenterology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari 70124, Italy
| | - Mariabeatrice Principi
- Gastroenterology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari 70124, Italy
| | - Michele Barone
- Gastroenterology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari 70124, Italy
| | - Enzo Ierardi
- Gastroenterology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari 70124, Italy
| | - Alfredo Di Leo
- Gastroenterology Section, Department of Emergency and Organ Transplantation, University of Bari, Bari 70124, Italy
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Camini FC, Costa DC. Silymarin: not just another antioxidant. J Basic Clin Physiol Pharmacol 2020; 31:/j/jbcpp.2020.31.issue-4/jbcpp-2019-0206/jbcpp-2019-0206.xml. [PMID: 32134732 DOI: 10.1515/jbcpp-2019-0206] [Citation(s) in RCA: 29] [Impact Index Per Article: 5.8] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/22/2019] [Accepted: 11/30/2019] [Indexed: 06/10/2023]
Abstract
Silymarin (Silybum marianum; SM), popularly known as milk thistle, is an extract that has been used for many centuries to treat liver diseases. In recent years, several studies have shown that SM is not only just another antioxidant but also a multifunctional compound that exhibits several beneficial properties for use in the treatment and prevention of different types of pathologies and disorders. This review aims at demonstrating the main protective activities of SM in diseases, such as cancer, diabetes, hepatitis, non-alcoholic fatty liver disease, alcoholic liver disease, hepatitis C virus, hepatitis B virus, metabolic syndrome, depression, cardiovascular diseases and thalassemia, in addition to its photoprotective activity in in vitro tests and preclinical studies. Its main functions include antioxidant and anti-inflammatory effects, and it acts as modulator of signaling pathways. It has been suggested that SM presents great multifunctional potential and is capable of achieving promising results in different types of research. However, caution is still needed regarding its indiscriminate use in humans as there are only a few clinical studies relating to the adequate dose and the actual efficacy of this extract in different types of diseases.
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Affiliation(s)
- Fernanda Caetano Camini
- Laboratory of Metabolic Biochemistry, Post-Graduate Program in Biological Sciences, Nucleus of Research in Biological Sciences, Federal University of Ouro Preto, Ouro Preto, Minas Gerais, Brazil
| | - Daniela Caldeira Costa
- Laboratory of Metabolic Biochemistry, Post-Graduate Program in Biological Sciences, Nucleus of Research in Biological Sciences, Federal University of Ouro Preto, Ouro Preto, Minas Gerais, Brazil
- Laboratory of Metabolic Biochemistry, Department of Biological Sciences, Federal University of Ouro Preto, Morro do Cruzeiro University Campus, Ouro Preto, Minas Gerais, Brazil
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Bektur Aykanat NE, Kacar S, Karakaya S, Sahinturk V. Silymarin suppresses HepG2 hepatocarcinoma cell progression through downregulation of Slit-2/Robo-1 pathway. Pharmacol Rep 2020; 72:199-207. [PMID: 32016841 DOI: 10.1007/s43440-019-00040-x] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/08/2019] [Revised: 09/19/2019] [Accepted: 10/11/2019] [Indexed: 02/06/2023]
Abstract
BACKGROUND 14 million people are diagnosed with new cancer and approximately 8 million people die from cancer every year. Hepatocellular carcinoma is the most common type of liver cancer and covers almost 5-6% of cancer deaths worldwide. Silybum marianum, a plant that contains silymarin, has been used traditionally in the treatment of liver diseases for centuries. The antioxidant, anti-inflammatory and anti-fibrotic anti-cancer properties of silymarin have been demonstrated in several studies in vivo and in vitro. The Slit/Robo signaling pathway plays a role in many processes such as neurogenesis, angiogenesis, cell proliferation, cell movement, cancer progression, cell invasion, migration and metastasis. In this study, we aimed to investigate the effects of silymarin on HepG2 Hepatocellular carcinoma cells on Slit-2/Robo-1 signaling pathway and CXCR-4 which plays a role in the metastasis process. METHODS HepG2 Hepatocellular carcinoma cells were used in the study. Different doses of silymarin's effect on HepG2 cells were observed by hematoxylin and eosin staining. Immunoblotting techniques were used to test the expression of Slit-2/Robo-1 and CXCR4 protein level. Immunocytochemistry was used to visualize the localization of Slit-2/Robo-1 and CXCR4 protein within the cells. RESULTS Silymarin caused apoptosis in HepG2 cells, decreased the level of CXCR-4 protein dose-dependently, and decreased the Slit-2/Robo-1 protein level at low doses and increased it at high doses. CONCLUSIONS Silymarin doses showed anti-carcinogenic, anti-metastatic and apoptotic effects in a dose-dependent manner on HepG2 cells through the Slit-2/Robo-1 pathway.
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Affiliation(s)
- Nuriye Ezgi Bektur Aykanat
- Department of Histology and Embryology, Faculty of Medicine, Eskisehir Osmangazi University, Eskisehir, Turkey.
| | - Sedat Kacar
- Department of Histology and Embryology, Faculty of Medicine, Eskisehir Osmangazi University, Eskisehir, Turkey
| | - Serife Karakaya
- Department of Histology and Embryology, Faculty of Medicine, Eskisehir Osmangazi University, Eskisehir, Turkey
| | - Varol Sahinturk
- Department of Histology and Embryology, Faculty of Medicine, Eskisehir Osmangazi University, Eskisehir, Turkey
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Inhibition of oxaliplatin-induced neurotoxicity by silymarin through increased expression of brain-derived neurotrophic factor and inhibition of p38-MAPK. Mol Cell Toxicol 2019. [DOI: 10.1007/s13273-019-0018-8] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/25/2022]
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Maruszewska A, Tarasiuk J. Antitumour effects of selected plant polyphenols, gallic acid and ellagic acid, on sensitive and multidrug-resistant leukaemia HL60 cells. Phytother Res 2019; 33:1208-1221. [PMID: 30838722 DOI: 10.1002/ptr.6317] [Citation(s) in RCA: 16] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/14/2018] [Revised: 01/18/2019] [Accepted: 01/24/2019] [Indexed: 12/30/2022]
Abstract
The aim of this study was to examine the antitumour effects of plant phenolic acids, gallic acid (GA) and ellagic acid (EA), on human promyelocytic leukaemia sensitive HL60 cell line and its resistant sublines exhibiting two MDR phenotypes: HL60/VINC (overexpressing P-glycoprotein) and HL60/MX2 (characterized by the presence of mutated α isoform of topoisomerase II). Both studied compounds exerted comparable cytotoxic activities towards sensitive HL60 cells and their MDR counterparts. It was also found that GA and EA modulated the cellular level of reactive oxygen species in a dose-dependent and time-dependent manner. Furthermore, it was demonstrated that GA (IC90 ) and EA (IC50 and IC90 ) significantly increased the percentage of sub-G1 subpopulation of all studied leukaemia cells causing oligonucleosomal DNA fragmentation. Both compounds used at IC90 triggered mainly the apoptotic death of these cells. However, GA had no effect on the activity of caspase-3 as well as caspase-8 in sensitive HL60 cells and their MDR counterparts. In contrast, EA provoked a significant activation of these caspases in all studied leukaemia cells. It was also found that lysosomes were not involved in triggering programmed death of sensitive HL60 and MDR cells by GA and EA.
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Affiliation(s)
- Agnieszka Maruszewska
- Department of Biochemistry, Faculty of Biology, University of Szczecin, 3c Felczaka St, Szczecin, 71-412, Poland.,Molecular Biology and Biotechnology Center, Faculty of Biology, University of Szczecin, 13 Wąska St, Szczecin, 71-415, Poland
| | - Jolanta Tarasiuk
- Department of Biochemistry, Faculty of Biology, University of Szczecin, 3c Felczaka St, Szczecin, 71-412, Poland.,Molecular Biology and Biotechnology Center, Faculty of Biology, University of Szczecin, 13 Wąska St, Szczecin, 71-415, Poland
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El-Marasy SA, Abd-Elsalam RM, Ahmed-Farid OA. Ameliorative Effect of Silymarin on Scopolamine-induced Dementia in Rats. Open Access Maced J Med Sci 2018; 6:1215-1224. [PMID: 30087724 PMCID: PMC6062269 DOI: 10.3889/oamjms.2018.257] [Citation(s) in RCA: 32] [Impact Index Per Article: 4.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/12/2018] [Revised: 05/23/2018] [Accepted: 05/25/2018] [Indexed: 12/14/2022] Open
Abstract
AIM: This study aims to elucidate the possible ameliorative effect of silymarin on scopolamine-induced dementia using the object recognition test (ORT) in rats. METHODS: The study was extended to demonstrate the role of cholinergic activity, oxidative stress, neuroinflammation, brain neurotransmitters and histopathological changes in the anti-amnestic effect of silymarin in demented rats. Wistar rats were pre-treated with silymarin (200, 400, 800 mg/kg) or donepezil (10 mg/kg) orally for 14 consecutive days. Dementia was induced after the last drug administration by a single intraperitoneal dose of scopolamine (16 mg/kg). Then behavioural, biochemical, histopathological, and immunohistochemical analyses were then performed. RESULTS: Rats pre-treated with silymarin counteracted scopolamine-induced non-spatial working memory impairment in the ORT and decreased acetylcholinesterase (AChE) activity, reduced malondialdehyde (MDA), elevated reduced glutathione (GSH), restored gamma-aminobutyric acid (GABA) and dopamine (DA) contents in the cortical and hippocampal brain homogenates. Silymarin reversed scopolamine-induced histopathological changes. Immunohistochemical analysis showed that silymarin mitigated protein expression of the glial fibrillary acidic protein (GFAP) and nuclear factor kappa-B (NF-κB) in the brain cortex and hippocampus. All these effects of silymarin were similar to that of the standard anti-amnestic drug, donepezil. CONCLUSION: This study reveals that the ameliorative effect of silymarin on scopolamine-induced dementia in rats using the ORT maybe in part mediated by, enhancement of cholinergic activity, anti-oxidant and anti-inflammatory activities as well as mitigation in brain neurotransmitters and histopathological changes.
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Affiliation(s)
| | - Reham M Abd-Elsalam
- Pathology Department, Faculty of Veterinary Medicine, Giza, Cairo University, Cairo, Egypt
| | - Omar A Ahmed-Farid
- Physiology Department, National Organization for Drug Control and Research, Giza, Egypt
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Effects of Silymarin-Loaded Nanoparticles on HT-29 Human Colon Cancer Cells. ACTA ACUST UNITED AC 2018; 54:medicina54010001. [PMID: 30344232 PMCID: PMC6037238 DOI: 10.3390/medicina54010001] [Citation(s) in RCA: 20] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/06/2017] [Revised: 02/26/2018] [Accepted: 02/26/2018] [Indexed: 12/26/2022]
Abstract
Background and objective: Previous studies have demonstrated the anti-cancer effects of silymarin (SLM). However, the low bioavailability of SLM has restricted its use. This study investigated the toxic effect of nanostructured SLM encapsulated in micelles (Nano-SLM) on the growth of the HT-29 human colon cancer cell line. Materials and methods: HT-29 cells were treated with 25 μM/mL of SLM or Nano-SLM for 48 h. MTT and colony formation assays were used to assess the cytotoxicity and proliferation of HT-29 cells, respectively. The cells were stained with annexin V/PI for assessment of apoptosis. Results: MTT assays revealed that Nano-SLM treatment was able to exert a more pronounced toxic effect on the HT-29 cells as compared to free SLM treatment (p < 0.01). In the Nano-SLM-treated cells, colony numbers were significantly reduced in comparison to the free SLM-treated cells (p < 0.01). Apoptotic and necrotic indexes of Nano-SLM-treated HT-29 cells were also significantly increased in comparison to those of the free SLM-treated cells (p < 0.01). The viability, proliferation and apoptosis of healthy cells (NIH-3T3 cells) were not changed in response to Nano-SLM or SLM. Conclusions: Our results indicate that Nano-SLM enhances the anti-cancer effects of SLM against human colon cancer cells.
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