军事医学2012，Vol.36Issue(12)：907-911,5.
HBx基因的真核表达及其对肝癌细胞系HepG2细胞表型的影响
The eukaryotic expression of hepatitis B virus X gene and its effect on the cellular phenotype of HepG2 cell line
摘要
Abstract
Objective To construct a eukaryotic expression vector for hepatitis B virus ( HBV) X protein and determine its effect on proliferation of hepatic carcinoma cell strain HepG 2. Methods A full-length sequence of HBx ORF was ampli -fied by PCR using plasmid( pCMV-HBx)as a template, and cloned into eukaryotic expression vector pcDNA3. 1-Flag. The constructed recombinant plasmid pcDNA3. 1-Flag-HBx was transfected to HepG2 cells. The transcription level of HBx mRNA by RT-PCR and expression level of HBx protein were deternined by Western blot . The cell proliferation rate via the 5-bromo-2'-deoxyuridine ( BrdU) and propidium iodide assay , while cell apoptosis by Annexin V-APC/7-AAD method. Results Restriction analysis and sequencing proved that recombinant eukaryotic expression vector pcDNA 3. 1-Flag-HBx was constructed correctly. The expression of HBx protein was proved in the HepG2 cells transfected with plasmid pcDNA3. 1-Flag-HBx. The cell proliferation ability of HepG2 cells transfected with plasmid pcDNA3. 1-Flag-HBx was increased compared with empty vector pcDNA3. 1-Flag. Meanwhile , the cell apoptosis ratio of HepG2 cells transfected with plasmid pcDNA3. 1 -Flag-HBx was lower than that with empty vector pcDNA 3. 1 -Flag. Conclusion The eukaryotic expres -sion vector for HBV X protein is successfully constructed , and the cell proliferation ability of HepG 2 cells transfected with plasmid pcDNA3. 1-Flag-HBx is increased compared with empty vector , while the cell apoptosis ratio of HepG2 cells transfected with plasmid pcDNA3. 1-Flag-HBx is lower than that with empty vector pcDNA3. 1-Flag. This research will help study the effect of HBx on the regulatory pathway of cell phenotype as well as the molecular mechanism of HBV -associated hepatocellular carcinoma ( HCC) caused by HBx.关键词
HBx/肝癌/真核细胞/细胞增殖/细胞凋亡Key words
hepatitis B virus x protein / hepatocellular carcinoma / eukaryotic cell/ cell proliferation / cell apoptosis分类
生物科学引用本文复制引用
刘玉玲,林巍然,郭英飞,杨冬,姜颖..HBx基因的真核表达及其对肝癌细胞系HepG2细胞表型的影响[J].军事医学,2012,36(12):907-911,5.基金项目
国家重点基础研究发展计划资助项目(2011CB910601) （2011CB910601）
国家自然科学基金资助项目(81000192) （81000192）
