• 2-D gel.
• LC-MS
• Prohibitin
• biomarker
• gastric cancer
• proteomics

• Humans
• Stomach Neoplasms/blood
• Stomach Neoplasms/diagnosis
• Proteomics
• Biomarkers, Tumor/blood
• Male
• India
• Female
• Middle Aged
• Prohibitins

• Histological detection
• Insect larvae
• Prohibitin
• Structural features
• Transcript levels
• Zabrotes subfasciatus

Stem cells maintain tissue homeostasis. We identified a factor, Yun, required for proliferation of normal and transformed intestinal stem cells in adult Drosophila. Yun acts as a scaffold to stabilize the Prohibitin (PHB) complex previously implicated in various cellular and developmental processes and diseases. The Yun/PHB complex acts downstream of EGFR/MAPK signaling and affects the levels of E2F1 to regulate intestinal stem cell proliferation. The role of the PHB complex in cell proliferation is evolutionarily conserved. Our results provide insight into the underlying mechanisms of how stem cell proliferation is properly controlled during tissue homeostasis and tumorigenesis.

Stem cells constantly divide and differentiate to maintain adult tissue homeostasis, and uncontrolled stem cell proliferation leads to severe diseases such as cancer. How stem cell proliferation is precisely controlled remains poorly understood. Here, from an RNA interference (RNAi) screen in adult Drosophila intestinal stem cells (ISCs), we identify a factor, Yun, required for proliferation of normal and transformed ISCs. Yun is mainly expressed in progenitors; our genetic and biochemical evidence suggest that it acts as a scaffold to stabilize the Prohibitin (PHB) complex previously implicated in various cellular and developmental processes and diseases. We demonstrate that the Yun/PHB complex is regulated by and acts downstream of EGFR/MAPK signaling. Importantly, the Yun/PHB complex interacts with and positively affects the levels of the transcription factor E2F1 to regulate ISC proliferation. In addition, we find that the role of the PHB complex in cell proliferation is evolutionarily conserved. Thus, our study uncovers a Yun/PHB-E2F1 regulatory axis in stem cell proliferation.

• Drosophila
• E2F1
• Prohibitin
• Yun
• intestinal stem cell

Prohibitin 1 (PHB1) is an evolutionarily conserved and ubiquitously expressed protein that stabilizes mitochondrial chaperone. Our previous studies showed that liver-specific Phb1 deficiency induced liver injuries and aggravated lipopolysaccharide (LPS)-induced innate immune responses. In this study, we performed RNA-sequencing (RNA-seq) analysis with liver tissues to investigate global gene expression among liver-specific Phb1^−/−, Phb1^+/−, and WT mice, focusing on the differentially expressed (DE) genes between Phb1^+/− and WT. When 78 DE genes were analyzed for biological functions, using ingenuity pathway analysis (IPA) tool, lipid metabolism-related genes, including insulin receptor (Insr), sterol regulatory element-binding transcription factor 1 (Srebf1), Srebf2, and SREBP cleavage-activating protein (Scap) appeared to be downregulated in liver-specific Phb1^+/− compared with WT. Diseases and biofunctions analyses conducted by IPA verified that hepatic system diseases, including liver fibrosis, liver hyperplasia/hyperproliferation, and liver necrosis/cell death, which may be caused by hepatotoxicity, were highly associated with liver-specific Phb1 deficiency in mice. Interestingly, of liver disease-related 5 DE genes between Phb1^+/− and WT, the mRNA expressions of forkhead box M1 (Foxm1) and TIMP inhibitor of metalloproteinase (Timp1) were matched with validation for RNA-seq in liver tissues and AML12 cells transfected with Phb1 siRNA. The results in this study provide additional insights into molecular mechanisms responsible for increasing susceptibility of liver injuries associated with hepatic Phb1.

• RNA-seq
• gene expression
• hepatotoxicity
• liver disease
• prohibitin 1

Low levels of reactive oxygen species (ROS) are crucial for maintaining cancer stem cells (CSCs) and their ability to resist therapy, but the ROS regulatory mechanisms in CSCs remains to be explored. Here, we discover that prohibitin (PHB) specifically regulates mitochondrial ROS production in glioma stem-like cells (GSCs) and facilitates GSC radiotherapeutic resistance. We find that PHB is upregulated in GSCs and is associated with malignant gliomas progression and poor prognosis. PHB binds to peroxiredoxin3 (PRDX3), a mitochondrion-specific peroxidase, and stabilizes PRDX3 protein through the ubiquitin-proteasome pathway. Knockout of PHB dramatically elevates ROS levels, thereby inhibiting GSC self-renewal. Importantly, deletion or pharmacological inhibition of PHB potently slows tumor growth and sensitizes tumors to radiotherapy, thus providing significant survival benefits in GSC-derived orthotopic tumors and glioblastoma patient-derived xenografts. These results reveal a selective role of PHB in mitochondrial ROS regulation in GSCs and suggest that targeting PHB improves radiotherapeutic efficacy in glioblastoma.

How ROS levels are regulated in cancer stem cells and their contribution to cancer resistance is currently not clear. Here, the authors show that prohibitin regulates mitochondrial ROS production stabilizing the peroxidase PRDX3 and this accounts for radiotherapy resistance in glioma stem-like cells.

The prohibitins (PHB) are SPFH domain-containing proteins found in the prokaryotes to eukaryotes. The plant PHBs are associated with a wide range of biological processes, including senescence, development, and responses to biotic and abiotic stresses. The PHB proteins are identified and characterized in the number of plant species, such as Arabidopsis, rice, maize, and soybean. However, no systematic identification of PHB proteins was performed in Solanum lycopersicum. In this study, we identified 16 PHB proteins in the tomato genome. The analysis of conserved motifs and gene structure validated the phylogenetic classification of tomato PHB proteins. It was observed that various members of tomato PHB proteins undergo purifying selection based on the Ka/Ks ratio and are targeted by four families of miRNAs. Moreover, SlPHB proteins displayed a very unique expression pattern in different plant parts including fruits at various development stages. It was found that SlPHBs processed various development-related and phytohormone responsive cis-regulatory elements in their promoter regions. Furthermore, the exogenous phytohormones treatments (Abscisic acid, indole-3-acetic acid, gibberellic acid, methyl jasmonate) salt and drought stresses induce the expression of SlPHB. Moreover, the subcellular localization assay revealed that SlPHB5 and SlPHB10 were located in the mitochondria. This study systematically summarized the general characterization of SlPHBs in the tomato genome and provides a foundation for the functional characterization of PHB genes in tomato and other plant species.

• Tomato
• expression
• phylogeny
• prohibitins
• stress
• synteny

• neuroprotection
• nitric oxide
• prohibitin
• protein S-nitrosylation

• Animals
• Cell Death/physiology
• Cells, Cultured
• Cyclic GMP/metabolism
• Enzyme Inhibitors/pharmacology
• Mice
• NG-Nitroarginine Methyl Ester/pharmacology
• Neurons/drug effects
• Neurons/metabolism
• Neuroprotection/drug effects
• Neuroprotection/physiology
• Nitric Oxide/metabolism
• Nitric Oxide Synthase/antagonists & inhibitors
• Prohibitins
• Reperfusion Injury/metabolism
• Repressor Proteins/metabolism
• Signal Transduction/drug effects
• Signal Transduction/physiology

• R01 NS034179 NINDS NIH HHS
• R01 NS067078 NINDS NIH HHS

FoxM1 amplification in human pancreatic cancer predicts poor prognosis and resistance to paclitaxel. Here, a novel role between FoxM1 (FoxM1b and FoxM1c) and Prohibitin1 (PHB1) in paclitaxel resistance has been identified. We adopted a bioinformatics approach to predict the potential effector of FoxM1. It specifically bound to the promoter of PHB1, and it enhanced PHB1 expression at transcriptional and post-transcriptional levels. FoxM1 contributed to the PHB1/C-RAF interaction and phosphorylation of ERK1/2 kinases, thus promoting paclitaxel resistance. Notably, FoxM1 conferred tumor cell resistance to paclitaxel, but knocking down PHB1 could sensitize pancreatic cancer cells to it. Besides, we identified that ABCA2 promoted paclitaxel resistance under the regulation of FoxM1/PHB1/RAF-MEK-ERK. Thiostrepton, an inhibitor of FoxM1, significantly decreased the expression of PHB1, p-ERK1/2, and ABCA2. It increased the influx of paclitaxel into the cell, and it attenuated FoxM1-mediated paclitaxel resistance in vitro and in vivo. Collectively, our findings defined PHB1 as an important downstream effector of FoxM1. It was regulated by FoxM1 to maintain phosphorylation of ERK1/2 in drug-resistant cells, and FoxM1 simultaneously enhanced the function of ABCA2, which collectively contributed to paclitaxel resistance. Targeting FoxM1 and its downstream effector PHB1 increased the sensitivity of pancreatic cells to paclitaxel treatment, providing potential therapeutic strategies for patients with paclitaxel resistance.

• ABCA2
• FoxM1
• MAPK signaling
• PHB1
• paclitaxel resistance
• pancreatic cancer
• thiostrepton

• Arabidopsis/cytology
• Arabidopsis/genetics
• Arabidopsis/growth & development
• Arabidopsis Proteins/genetics
• Arabidopsis Proteins/metabolism
• Arabidopsis Proteins/physiology
• Cell Cycle/genetics
• Cell Proliferation/genetics
• DNA Damage
• Gene Expression Regulation, Plant
• Genomic Instability
• Meristem/cytology
• Meristem/genetics
• Meristem/growth & development
• Minichromosome Maintenance Complex Component 2/genetics
• Minichromosome Maintenance Complex Component 2/metabolism
• Minichromosome Maintenance Complex Component 2/physiology
• Minichromosome Maintenance Proteins/genetics
• Minichromosome Maintenance Proteins/metabolism
• Minichromosome Maintenance Proteins/physiology
• Prohibitins
• Reactive Oxygen Species/metabolism
• Repressor Proteins/genetics
• Repressor Proteins/metabolism
• Repressor Proteins/physiology

• National Natural Science Foundation of China (NSFC)

To investigate the functional role and underlying molecular mechanism of miR-29a in hepatitis B virus (HBV) expression and replication.

The levels of miR-29a and SMARCE1 in HBV-infected HepG2.2.15 cells were measured by quantitative real-time PCR and western blot analysis. HBV DNA replication was measured by quantitative PCR and Southern blot analysis. The relative levels of hepatitis B surface antigen and hepatitis B e antigen were detected by enzyme-linked immunosorbent assay. The Cell Counting Kit-8 (CCK-8) was used to detect the viability of HepG2.2.15 cells. The relationship between miR-29a and SMARCE1 were identified by target prediction and luciferase reporter analysis.

miR-29a promoted HBV replication and expression, while SMARCE1 repressed HBV replication and expression. Cell viability detection indicated that miR-29a transfection had no adverse effect on the host cells. Moreover, SMARCE1 was identified and validated to be a functional target of miR-29a. Furthermore, restored expression of SMARCE1 could relieve the increased HBV replication and expression caused by miR-29a overexpression.

miR-29a promotes HBV replication and expression through regulating SMARCE1. As a potential regulator of HBV replication and expression, miR-29a could be a promising therapeutic target for patients with HBV infection.

• miR-29a
• SMARCE1
• Hepatitis B surface antigen
• Hepatitis B virus replication
• Hepatitis B e antigen

• Blotting, Western
• Carcinoma, Hepatocellular/genetics
• Carcinoma, Hepatocellular/virology
• Cell Survival
• Chromosomal Proteins, Non-Histone/genetics
• Chromosomal Proteins, Non-Histone/metabolism
• DNA Replication
• DNA, Viral/isolation & purification
• DNA, Viral/metabolism
• DNA-Binding Proteins/genetics
• DNA-Binding Proteins/metabolism
• Enzyme-Linked Immunosorbent Assay
• Gene Expression Regulation, Neoplastic
• Gene Expression Regulation, Viral
• Hep G2 Cells
• Hepatitis B/genetics
• Hepatitis B Surface Antigens/isolation & purification
• Hepatitis B Surface Antigens/metabolism
• Hepatitis B e Antigens/isolation & purification
• Hepatitis B e Antigens/metabolism
• Hepatitis B virus/genetics
• Humans
• Liver Neoplasms/genetics
• Liver Neoplasms/virology
• MicroRNAs/metabolism
• Real-Time Polymerase Chain Reaction
• Up-Regulation
• Virus Replication/genetics

• Adipocyte
• adipose-immune crosstalk
• immune cell
• mitochondria
• pleiotropic protein
• sex dimorphic

Natural compounds from soft corals have been increasingly used for their antitumor therapeutic properties. This study examined 11-epi-sinulariolide acetate (11-epi-SA), an active compound isolated from the cultured soft coral Sinularia flexibilis, to determine its potential antitumor effect on four hepatocellular carcinoma cell lines. Cell viability was investigated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and the results demonstrated that 11-epi-SA treatment showed more cytotoxic effect toward HA22T cells. Protein profiling of the 11-epi-SA-treated HA22T cells revealed substantial protein alterations associated with stress response and protein synthesis and folding, suggesting that the mitochondria and endoplasmic reticulum (ER) play roles in 11-epi-SA-initiated apoptosis. Moreover, 11-epi-SA activated caspase-dependent apoptotic cell death, suggesting that mitochondria-related apoptosis genes were involved in programmed cell death. The unfolded protein response signaling pathway-related proteins were also activated on 11-epi-SA treatment, and these changes were accompanied by the upregulated expression of growth arrest and DNA damage-inducible protein (GADD153) and CCAAT/enhancer binding protein (C/EBP) homologous protein (CHOP), the genes encoding transcription factors associated with growth arrest and apoptosis under prolonged ER stress. Two inhibitors, namely salubrinal (Sal) and SP600125, partially abrogated 11-epi-SA-related cell death, implying that the protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK)–activating transcription factor (ATF) 6–CHOP or the inositol-requiring enzyme 1 alpha (IRE1α)–c-Jun N-terminal kinase (JNK)–cJun signal pathway was activated after 11-epi-SA treatment. In general, these results suggest that 11-epi-SA exerts cytotoxic effects on HA22T cells through mitochondrial dysfunction and ER stress cell death pathways.

• 11-epi-sinulariolide acetate
• ER stress
• antitumor
• hepatocellular carcinoma
• mitochondrial dysfunction

• Animals
• Anthozoa/chemistry
• Anthracenes/administration & dosage
• Apoptosis/drug effects
• Carcinoma, Hepatocellular/drug therapy
• Carcinoma, Hepatocellular/genetics
• Carcinoma, Hepatocellular/pathology
• Cell Proliferation/drug effects
• Cell Survival/drug effects
• Cinnamates/administration & dosage
• Diterpenes/administration & dosage
• Diterpenes/chemical synthesis
• Diterpenes/chemistry
• Endoplasmic Reticulum Stress/drug effects
• Gene Expression Regulation, Neoplastic/drug effects
• Humans
• JNK Mitogen-Activated Protein Kinases/biosynthesis
• JNK Mitogen-Activated Protein Kinases/genetics
• Liver Neoplasms/drug therapy
• Liver Neoplasms/genetics
• Liver Neoplasms/pathology
• Mitochondria/drug effects
• Mitochondria/pathology
• Signal Transduction/drug effects
• Thiourea/administration & dosage
• Thiourea/analogs & derivatives
• Transcription Factor CHOP/biosynthesis
• Transcription Factor CHOP/genetics
• Unfolded Protein Response/drug effects

• Bacteria-containing vacuole
• Listeria
• Prohibitin
• Rab
• Salmonella
• Xenophagy

• Dendritic Cells/immunology
• Dendritic Cells/microbiology
• Humans
• Listeria monocytogenes/immunology
• Lysosomes/microbiology
• Macrophages/immunology
• Macrophages/microbiology
• Phagosomes/microbiology
• Salmonella/immunology
• rab GTP-Binding Proteins/metabolism

• advanced ovarian cancer
• apoptotic pathways
• biomarkers
• chemotherapy response
• gel-based proteomics

• RtxA1 toxin
• V. vulnificus
• prohibitin 1

• Cancer
• Chromatin
• Chromatin remodeling
• Epigenetics
• Nucleosome

• Adenosine Triphosphate/pharmacology
• Animals
• Chromatin Assembly and Disassembly/drug effects
• Histones/metabolism
• Humans
• Molecular Targeted Therapy/methods
• Molecular Targeted Therapy/trends
• Multiprotein Complexes/physiology
• Neoplasms/genetics
• Neoplasms/pathology
• Neoplasms/therapy
• Nucleosomes/metabolism
• Transcription Factors/metabolism

• P30 CA016059 NCI NIH HHS

Pollen tubes are the vehicle for sperm cell delivery to the embryo sac during fertilisation of Angiosperms. They provide an intriguing model for unravelling mechanisms of growing to extremes. The asymmetric distribution of lipids and proteins in the pollen tube plasma membrane modulates ion fluxes and actin dynamics and is maintained by a delicate equilibrium between exocytosis and endocytosis. The structural constraints regulating polarised secretion and asymmetric protein distribution on the plasma membrane are mostly unknown. To address this problem, we investigated whether ordered membrane microdomains, namely membrane rafts, might contribute to sperm cell delivery. Detergent insoluble membranes, rich in sterols and sphingolipids, were isolated from tobacco pollen tubes. MALDI TOF/MS analysis revealed that actin, prohibitins and proteins involved in methylation reactions and in phosphoinositide pattern regulation are specifically present in pollen tube detergent insoluble membranes. Tubulins, voltage-dependent anion channels and proteins involved in membrane trafficking and signalling were also present. This paper reports the first evidence of membrane rafts in Angiosperm pollen tubes, opening new perspectives on the coordination of signal transduction, cytoskeleton dynamics and polarised secretion.

• Membrane microdomains
• Nicotiana tabacum (L.)
• Pollen tube

• Human chang liver cells
• Lamprey
• Oxidative stress
• Prohibitin
• Translocation

• Amino Acid Sequence
• Animals
• Cloning, Molecular
• DNA, Complementary/genetics
• DNA, Complementary/metabolism
• Female
• Fish Proteins/genetics
• Fish Proteins/metabolism
• Gene Expression Regulation/drug effects
• Hydrogen Peroxide/pharmacology
• Lampreys/genetics
• Lampreys/immunology
• Lampreys/metabolism
• Male
• Molecular Sequence Data
• Organ Specificity
• Oxidative Stress/drug effects
• Phylogeny
• Prohibitins
• RNA, Messenger/genetics
• RNA, Messenger/metabolism
• Real-Time Polymerase Chain Reaction/veterinary
• Repressor Proteins/genetics
• Repressor Proteins/metabolism
• Sequence Alignment/veterinary

• wt1
• basp1
• prohibitin
• transcription

• Cell Line, Tumor
• Cell Nucleus/metabolism
• Chromatin Assembly and Disassembly
• Gene Expression Regulation, Neoplastic
• HeLa Cells
• Humans
• K562 Cells
• MCF-7 Cells
• Membrane Proteins/metabolism
• Nerve Tissue Proteins/metabolism
• Phospholipids/metabolism
• Prohibitins
• Promoter Regions, Genetic
• Repressor Proteins/metabolism
• Transcription, Genetic
• WT1 Proteins/genetics
• WT1 Proteins/metabolism

• A6630 Cancer Research UK
• R01 GM098609 NIGMS NIH HHS
• 1R01GM098609 NIGMS NIH HHS
• C1356/A6630 Cancer Research UK

Spermatozoa are more efficient at supporting normal embryonic development than spermatids, their immature, immediate precursors. This suggests that the sperm acquires the ability to support embryonic development during spermiogenesis (spermatid to sperm maturation). Here, using Xenopus laevis as a model organism, we performed 2-D Fluorescence Difference Gel Electrophoresis (2D-DIGE) and mass spectrometry analysis of differentially expressed proteins between sperm and spermatids in order to identify factors that could be responsible for the efficiency of the sperm to support embryonic development. Furthermore, benefiting from the availability of egg extracts in Xenopus, we also tested whether the chromatin of sperm could attract different egg factors compared to the chromatin of spermatids. Our analysis identified: (1) several proteins which were present exclusively in sperm; but not in spermatid nuclei and (2) numerous egg proteins binding to the sperm (but not to the spermatid chromatin) after incubation in egg extracts. Amongst these factors we identified many chromatin-associated proteins and transcriptional repressors. Presence of transcriptional repressors binding specifically to sperm chromatin could suggest its preparation for the early embryonic cell cycles, during which no transcription is observed and suggests that sperm chromatin has a unique protein composition, which facilitates the recruitment of egg chromatin remodelling factors. It is therefore likely that the acquisition of these sperm-specific factors during spermiogenesis makes the sperm chromatin suitable to interact with the maternal factors and, as a consequence, to support efficient embryonic development.

• Animals
• Chromatin/metabolism
• Chromatin Assembly and Disassembly
• Egg Proteins/metabolism
• Electrophoresis, Polyacrylamide Gel
• Female
• Immunoblotting
• Male
• Mass Spectrometry
• Nuclear Proteins/isolation & purification
• Nuclear Proteins/metabolism
• Protein Binding
• Protein Interaction Mapping
• Protein Isoforms
• Sperm-Ovum Interactions
• Spermatids/metabolism
• Spermatogenesis/physiology
• Spermatozoa/metabolism
• Tissue Extracts
• Xenopus Proteins/isolation & purification
• Xenopus Proteins/metabolism
• Xenopus laevis/metabolism

• WT089613 Wellcome Trust
• 092096 Wellcome Trust
• MR/K011022/1 Medical Research Council
• C6946/A14492 Cancer Research UK
• 101050 Wellcome Trust
• WT101050/Z/13/Z Wellcome Trust
• BBS/B/14647 Biotechnology and Biological Sciences Research Council

Paclitaxel is a drug within one of the most promising classes of anticancer agents. Unfortunately, clinical success of this drug has been limited by the insurgence of cellular resistance. To address this, Paclitaxel resistance was modeled in an in vitro system using estrogen treated prostate cancer cells. This study demonstrates that emerging resistance to clinically relevant doses of Paclitaxel is associated with 17-β-estradiol (E2) treatment in PC-3 cells, but not in LNCaP cells. We found that small interfering RNA mediated knockdown of ERα lead to a decrease in E2 induced Paclitaxel resistance in androgen-independent cells. We also showed that ERα mediated the effects of estrogen, thereby suppressing androgen-independent cell proliferation and mediating Paclitaxel resistance. Furthermore, E2 promoted Prohibitin (PHB) mitochondrial-nucleus translocation via directly mediation of ERα, leading to an inhibition of cellular proliferation by PHB. Additionally, restoration of Paclitaxel sensitivity by ERα knockdown could be overcome by PHB overexpression and, conversely, PHB knockdown decreased E2 induced Paclitaxel resistance. These findings demonstrate that PHB lies downstream of ERα and mediates estrogen-dependent Paclitaxel resistance signaling cascades.

• Active Transport, Cell Nucleus/drug effects
• Antineoplastic Agents/pharmacology
• Cell Death/drug effects
• Cell Line, Tumor
• Cell Nucleus/drug effects
• Cell Nucleus/metabolism
• Cell Proliferation/drug effects
• Drug Resistance, Neoplasm
• Estrogen Receptor alpha/genetics
• Estrogen Receptor alpha/metabolism
• Estrogens/pharmacology
• Gene Expression Regulation, Neoplastic/drug effects
• Humans
• Male
• Mitochondria/drug effects
• Mitochondria/metabolism
• Paclitaxel/pharmacology
• Prohibitins
• Prostatic Neoplasms/pathology
• Repressor Proteins/metabolism

• Antigens, Nuclear/genetics
• Antigens, Nuclear/metabolism
• Chromatography, Liquid
• DNA/genetics
• DNA/metabolism
• DNA Damage
• DNA Repair
• DNA-Binding Proteins/genetics
• DNA-Binding Proteins/metabolism
• Dual Specificity Phosphatase 3/genetics
• Dual Specificity Phosphatase 3/metabolism
• Gamma Rays
• Gene Expression Regulation
• HeLa Cells
• Heterogeneous-Nuclear Ribonucleoprotein Group C/genetics
• Heterogeneous-Nuclear Ribonucleoprotein Group C/metabolism
• Humans
• Immunoprecipitation
• Ku Autoantigen
• Nuclear Proteins/genetics
• Nuclear Proteins/metabolism
• Nucleophosmin
• Phosphoproteins/genetics
• Phosphoproteins/metabolism
• Protein Binding
• Protein Interaction Mapping
• RNA-Binding Proteins/genetics
• RNA-Binding Proteins/metabolism
• Recombinant Fusion Proteins/genetics
• Recombinant Fusion Proteins/metabolism
• Signal Transduction
• Tandem Mass Spectrometry
• Telomerase/genetics
• Telomerase/metabolism
• Ultraviolet Rays
• Nucleolin

• ERK
• NAD(P)H quinone oxidoreductase-1
• Nrf2
• activator protein-1
• heme oxygenase-1
• inflammatory bowel disease
• oxidative stress
• prohibitin

Identifying novel tumor biomarkers to develop more effective diagnostic and therapeutic strategies for patients with ACC is urgently needed. The aim of the study was to compare the proteomic profiles between adrenocortical carcinomas (ACC) and normal adrenocortical tissues in order to identify novel potential biomarkers for ACC.

The protein samples from 12 ACC tissues and their paired adjacent normal adrenocortical tissues were profiled with two-dimensional electrophoresis; and differentially expressed proteins were identified by mass spectrometry. Expression patterns of three differently expressed proteins calreticulin, prohibitin and HSP60 in ACC, adrenocortical adenomas (ACA) and normal adrenocortical tissues were further validated by immunohistochemistry.

In our proteomic study, we identified 20 up-regulated and 9 down-regulated proteins in ACC tissues compared with paired normal controls. Most of the up-regulated proteins were focused in protein binding and oxidoreductase activity in Gene Ontology (GO) molecular function classification. By immunohistochemistry, two biomarkers calreticulin and prohibitin were validated to be overexpressed in ACC compared with adrenocortical adenomas (ACA) and normal tissues, but also calreticulin overexpression was significantly associated with tumor stages of ACC.

For the first time, calreticulin and prohibitin were identified to be novel candidate biomarkers for ACC, and their roles during ACC carcinogenesis and clinical significance deserves further investigation.

The virtual slides for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/1897372598927465

• Cell Count
• Cell Cycle Proteins/genetics
• Cell Cycle Proteins/metabolism
• Cell Nucleus/genetics
• Cell Nucleus/metabolism
• Centromere/genetics
• Centromere/metabolism
• Chromatids/genetics
• Chromatids/metabolism
• Chromatin/genetics
• Chromatin/metabolism
• Chromatin Assembly and Disassembly
• Chromosomal Proteins, Non-Histone/genetics
• Chromosomal Proteins, Non-Histone/metabolism
• DNA-Binding Proteins
• Fluorescent Dyes/metabolism
• G2 Phase Cell Cycle Checkpoints
• HeLa Cells
• Humans
• Mitochondria/genetics
• Mitochondria/metabolism
• Mitosis
• Nuclear Proteins/genetics
• Nuclear Proteins/metabolism
• Phosphoproteins/genetics
• Phosphoproteins/metabolism
• Prohibitins
• Protein Binding
• Protein Interaction Mapping/methods
• Protein Structure, Tertiary
• RNA Interference
• Repressor Proteins/genetics
• Repressor Proteins/metabolism
• Cohesins

• Animals
• Digestive System Diseases/metabolism
• Humans
• Kidney Diseases/metabolism
• Liver Diseases/metabolism
• Lung Diseases/metabolism
• Neoplasms/metabolism
• Prohibitins
• Repressor Proteins/metabolism
• Signal Transduction

The E2F6 protein functions as an Rb-independent repressor of gene transcription. We have previously provided evidence suggesting a role for E2F6 in repression of E2F-responsive genes at S phase. Here, we have identified BRG1, the ATPase subunit of the SWI/SNF chromatin-remodeling complex, as an E2F6 interacting protein. Immunoprecipitation experiments demonstrate that BRG1 binds specifically to E2F6 and E2F4 but not the activator E2Fs. E2F6 was also able to interact with BAF155, a BRG1-associated factor, in the SWI/SNF complex. Chromatin immunoprecipitation assays demonstrate the binding of BRG1 coincident with E2F6 on G1/S gene promoters during S phase. Collectively, our studies suggest that E2F6 may recruit BRG1 in transcriptional regulation of genes important for G1/S phase transition of the cell cycle.

• Chromatin Immunoprecipitation
• DNA Helicases/metabolism
• E2F1 Transcription Factor/genetics
• E2F4 Transcription Factor/metabolism
• E2F6 Transcription Factor/metabolism
• Gene Expression Regulation
• Genes, Dominant
• Humans
• Nuclear Proteins/metabolism
• Promoter Regions, Genetic
• Protein Binding/genetics
• Transcription Factors/metabolism
• Transcription, Genetic
• Two-Hybrid System Techniques

• R01-CA104663 NCI NIH HHS
• R01 CA106520 NCI NIH HHS
• U54 CA112952 NCI NIH HHS
• R01 CA104663 NCI NIH HHS
• U54-CA112952 NCI NIH HHS

Autophagy is an adaptive response to extracellular and intracellular stress by which cytoplasmic components and organelles, including damaged mitochondria, are degraded to promote cell survival and restore cell homeostasis. Certain genes involved in autophagy confer susceptibility to Crohn's disease. Reactive oxygen species and pro-inflammatory cytokines such as tumor necrosis factor α (TNFα), both of which are increased during active inflammatory bowel disease, promote cellular injury and autophagy via mitochondrial damage. Prohibitin (PHB), which plays a role in maintaining normal mitochondrial respiratory function, is decreased during active inflammatory bowel disease. Restoration of colonic epithelial PHB expression protects mice from experimental colitis and combats oxidative stress. In this study, we investigated the potential role of PHB in modulating mitochondrial stress-related autophagy in intestinal epithelial cells.

We measured autophagy activation in response to knockdown of PHB expression by RNA interference in Caco2-BBE and HCT116 WT and p53 null cells. The effect of exogenous PHB expression on TNFα- and IFNγ-induced autophagy was assessed. Autophagy was inhibited using Bafilomycin A₁ or siATG16L1 during PHB knockdown and the affect on intracellular oxidative stress, mitochondrial membrane potential, and cell viability were determined. The requirement of intracellular ROS in siPHB-induced autophagy was assessed using the ROS scavenger N-acetyl-L-cysteine.

TNFα and IFNγ-induced autophagy inversely correlated with PHB protein expression. Exogenous PHB expression reduced basal autophagy and TNFα-induced autophagy. Gene silencing of PHB in epithelial cells induces mitochondrial autophagy via increased intracellular ROS. Inhibition of autophagy during PHB knockdown exacerbates mitochondrial depolarization and reduces cell viability.

Decreased PHB levels coupled with dysfunctional autophagy renders intestinal epithelial cells susceptible to mitochondrial damage and cytotoxicity. Repletion of PHB may represent a therapeutic approach to combat oxidant and cytokine-induced mitochondrial damage in diseases such as inflammatory bowel disease.

• prohibitins
• phb
• phb2
• repressor of estrogen receptor action
• rea
• mitochondria
• nucleus
• review

• Androgens/physiology
• Apoptosis
• Cell Differentiation
• Female
• Gonadotropins/physiology
• Humans
• Ovarian Follicle/growth & development
• Phosphorylation
• Prohibitins
• Repressor Proteins/metabolism
• Repressor Proteins/physiology
• Subcellular Fractions/metabolism

• R01 HD057235-02 NICHD NIH HHS
• G12 RR003034 NCRR NIH HHS
• C06 RR018386 NCRR NIH HHS
• R01 HD057235 NICHD NIH HHS
• C06 RR18386 NCRR NIH HHS
• HD41749 NICHD NIH HHS
• G12-RR03034 NCRR NIH HHS
• 1R01HD057235 NICHD NIH HHS
• S21 MD000101 NIMHD NIH HHS
• U54 HD041749 NICHD NIH HHS

• prohibitin
• therapeutic
• cancer
• inflammation
• obesity
• diabetes

• Diabetes Mellitus, Type 2/genetics
• Diabetes Mellitus, Type 2/physiopathology
• Gene Expression Regulation
• Humans
• Inflammatory Bowel Diseases/genetics
• Inflammatory Bowel Diseases/physiopathology
• Mitochondria/metabolism
• Models, Biological
• Neoplasms/genetics
• Neoplasms/physiopathology
• Obesity/genetics
• Obesity/physiopathology
• Oxidative Stress/physiology
• Prohibitins
• Repressor Proteins/metabolism
• Repressor Proteins/physiology

• K01 DK085222 NIDDK NIH HHS
• R01 DK064711 NIDDK NIH HHS
• K01-DK085222 NIDDK NIH HHS
• R01-DK06411 NIDDK NIH HHS