医学分子生物学杂志2012，Vol.9Issue(1)：42-48,7.DOI:10.3870/j.issn.1672-8009.2012.01.009
丙型肝炎病毒E1蛋白活化MAPK/ERK信号通路促进肝癌细胞增殖
Hepatitis C Virus E1 Protein Promotes Hepatoma Cell Growth Through MAPK/Erk Signaling Pathway
摘要
Abstract
Objective To investigate the biological function of El protein encoded by Hepatitis C virus (HCV) .Methods SMMC-7721 cells were infected with adenovirus Ad-El, Ad-E2, Ad-NS3, Ad-NS5a and Ad-NS5b, which encodes El, E2, NS3, NS5a and NS5b protein, respectively. Over-expression of the target gene was identified by Western blot or RT-PCR methods. Cells were infected with high titer of adenovirus. Biological function induced by different viral protein was determined by cell proliferation assays. The cell growth and cell cycle were analyzed by MTS, Crystal Violet assay and flow cytometry. The transcriptional factor and target genes of MAPK/ ERK signal pathway were determined by Western blot or RT-PCR methods. Results RNA transcriptproducts of El, E2, NS3, NS5a and NS5b were identified by RT-PCR method respectively. El Protein expression was verified by Western blot analysis. Cell growth curve, MTS and cell viability assay indicated that over-expression of HCV El alone resulted in an increased proliferation rate in SMMC-7721 cells. Cell cycle analysis showed that the S-phase distribution in cells transduced with El was 34. 38%, much higher than that of GFP control (27.32%) (P<0. 05) .Moreover, El moderately up-regulated mRNA expression levels of several transcriptional factors downstream of MAPK/ERK signal pathway. Conclusion The growth effect of overexpressed HCV El Protein in vitro on SMMC-7721 cells was partially correlated with MAPK/ERK signal pathway.关键词
丙型肝炎病毒/E1蛋白/丝裂原活化蛋白激酶/细胞增殖Key words
hepatitis C virus/ El protein/ mitogen-activated protein kinases/ cell proliferation分类
医药卫生引用本文复制引用
陈庆美,刘娇,单晓亮,周帆,权会琴,陈林林,唐霓..丙型肝炎病毒E1蛋白活化MAPK/ERK信号通路促进肝癌细胞增殖[J].医学分子生物学杂志,2012,9(1):42-48,7.基金项目
国家自然科学基金(No.30972586,31171307) （No.30972586,31171307）
重庆市自然科学基金重点项目(No.CSTC 2009BA5036) （No.CSTC 2009BA5036）
