北京大学学报（医学版）2012，Vol.44Issue(6)：932-936,5.DOI:10.3969/j.issn.1671-167X.2012.06.026
HBx诱导p16INK4a高甲基化在Chang细胞株恶性转化倾向中的作用
Hepatitis B virus X protein induces malignant transformation tendency of Chang liver cells involving p16INK4a hypermethylation
摘要
Abstract
Objective: To study the relationship between hepatitis B virus X (HBx) protein and DNA methylation of pl6 INK4a and the role of HBx in the carcinogenesis of hepatocellular carcinoma. Methods; Eukaryonic expression vectors pcDNA3. lB-HBx and pcDNA3. IB were transduced into Chang liver cells by using Lipofectamine 2000 to establish the Chang-HBx liver cell line ( HBx expression) and Chang-vector liver cell line (non-HBx expression). RT-PCR and Western blot were used to test the expression of pl6INK4 in the two cell lines. The level of pl6INK4a promoter methylation was tested by methylation specific PCR ( MSP) . The proliferation curves were drew by CCK-8 , and S phase in cell cycle and apoptosis were observed by flow cytometry. Results; Hypermethylation of pl6 can be mediated by HBx, which decreases the expression of mRNA and protein of pl6. Chang-HBx cells grow faster. Chang-HBx cells have much higher S-phase population (28. 96% vs. 21. 53% , P <0. 001 ; 28. 96% vs. 21. 5% , P <0. 001) and lower apoptosis rate (2. 71% vs. 3. 69% , P <0. 001 ; 2. 71% vs. 3. 36% , P <0. 001) than Chang-vector cells and Chang cells respectively. Conclusion; pl6 "expression was repressed by HBx protein via DNA methylation of pl6INK4a, which can induce the malignant transformation tendency of Chang cells.关键词
癌,肝细胞/乙型肝炎病毒X蛋白/基因,p16/甲基化Key words
Carcinoma, hepatocelluar/ Hepatitis B virus X protein/ Genes, p16/ Methylation分类
医药卫生引用本文复制引用
吕文平,周开伦,高彬,陈永亮,向昕,苏明,董家鸿..HBx诱导p16INK4a高甲基化在Chang细胞株恶性转化倾向中的作用[J].北京大学学报（医学版）,2012,44(6):932-936,5.基金项目
国家自然科学基金(30772493)资助 （30772493）
