Document Detail


Bcl-2 overexpression in hepatic stellate cell line CFSC-2G, induces a pro-fibrotic state.
MedLine Citation:
PMID:  20594261     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND AND AIM: Development of hepatic fibrosis is a complex process that involves oxidative stress (OS) and an altered balance between pro- and anti-apoptotic molecules. Since Bcl-2 overexpression preserves viability against OS, our objective was to address the effect of Bcl-2 overexpression in the hepatic stellate cells (HSC) cell-line CFSC-2G under acetaldehyde and H(2)O(2) challenge, and explore if it protects these cells against OS, induces replicative senescence and/or modify extracellular matrix (ECM) remodeling potential. METHODS: To induce Bcl-2 overexpression, HSC cell line CFSC-2G was transfected by lipofection technique. Green fluorescent protein-only CFSC-2G cells were used as a control. Cell survival after H(2)O(2) treatment and total protein oxidation were assessed. To determine cell cycle arrest, proliferation-rate, DNA synthesis and senescence were assessed. Matrix metalloproteinases (MMP), tissue-inhibitor of MMP (TIMP), transglutaminases (TG) and smooth muscle a-actin (alpha-SMA) were evaluated by western blot in response to acetaldehyde treatment as markers of ECM remodeling capacity in addition to transforming growth factor-beta (TGF-beta) mRNA. RESULTS: Cells overexpressing Bcl-2 survived approximately 20% more than control cells when exposed to H(2)O(2) and approximately 35% proteins were protected from oxidation, but Bcl-2 did not slow proliferation or induced senescence. Bcl-2 overexpression did not change alpha-SMA levels, but it increased TIMP-1 (55%), tissue transglutaminases (tTG) (25%) and TGF-beta mRNA (49%), when exposed to acetaldehyde, while MMP-13 content decreased (47%). CONCLUSIONS: Bcl-2 overexpression protected HSC against oxidative stress but it did not induce replicative senescence. It increased TIMP-1, tTG and TGF-beta mRNA levels and decreased MMP-13 content, suggesting that Bcl-2 overexpression may play a key role in the progression of fibrosis in chronic liver diseases.
Authors:
Viridiana Y González-Puertos; Elizabeth Hernández-Pérez; Natalia Nuño-Lámbarri; José L Ventura-Gallegos; Norma E López-Diázguerrero; Guillermo Robles-Díaz; María C Gutiérrez-Ruiz; Mina Konigsberg
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of gastroenterology and hepatology     Volume:  25     ISSN:  1440-1746     ISO Abbreviation:  J. Gastroenterol. Hepatol.     Publication Date:  2010 Jul 
Date Detail:
Created Date:  2010-07-02     Completed Date:  2010-10-18     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8607909     Medline TA:  J Gastroenterol Hepatol     Country:  Australia    
Other Details:
Languages:  eng     Pagination:  1306-14     Citation Subset:  IM    
Affiliation:
Health and Sciences Department, Biological and Health Sciences Division, Metropolitan Independent University, México, DF, México.
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MeSH Terms
Descriptor/Qualifier:
Acetaldehyde / pharmacology
Actins / metabolism
Animals
Cell Aging
Cell Line
Cell Proliferation
DNA Replication
Dose-Response Relationship, Drug
Extracellular Matrix / metabolism
GTP-Binding Proteins / metabolism
Hepatic Stellate Cells / drug effects,  metabolism*,  pathology
Humans
Hydrogen Peroxide / pharmacology
Liver Cirrhosis / genetics,  metabolism*,  pathology
Matrix Metalloproteinase 13 / metabolism
Oxidants / pharmacology
Oxidative Stress
Proto-Oncogene Proteins c-bcl-2 / genetics,  metabolism*
RNA, Messenger / metabolism
Rats
Recombinant Fusion Proteins / metabolism
Time Factors
Tissue Inhibitor of Metalloproteinase-1 / metabolism
Transfection
Transforming Growth Factor beta / genetics
Transglutaminases / metabolism
Up-Regulation
Chemical
Reg. No./Substance:
0/Actins; 0/Oxidants; 0/Proto-Oncogene Proteins c-bcl-2; 0/RNA, Messenger; 0/Recombinant Fusion Proteins; 0/Tissue Inhibitor of Metalloproteinase-1; 0/Transforming Growth Factor beta; 0/smooth muscle actin, rat; 75-07-0/Acetaldehyde; 7722-84-1/Hydrogen Peroxide; EC 2.3.2.-/transglutaminase 2; EC 2.3.2.13/Transglutaminases; EC 3.4.24.-/Matrix Metalloproteinase 13; EC 3.4.24.-/Mmp13 protein, rat; EC 3.6.1.-/GTP-Binding Proteins

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