目的研究丙型肝炎病毒(hepatitis C virus,HCV)基因1b型F蛋白对HepG2细胞中Bcl-2、Bax表达的影响。方法应用聚合酶链式反应(polymerase chain reaction,PCR)技术扩增HCV 1b型的F基因,构建pEGFP-C3-F、pEGFP-C3-C真核表达载体,分别将pEGFP-C3-F、pEGFP-C3-C、pEGFP-C3(阴性对照)瞬时转染HepG2细胞,48h后抽细胞总蛋白质,蛋白质印迹法(Western blot)检测Bcl-2、Bax表达的变化。结果转染F基因的HepG2中Bcl-2表达水平略高于未转染的细胞,但Bax表达水平显著高于未转染的细胞。结论 HCV F蛋白具有调节原癌基因Bcl-2和抑癌基因Bax的特性,可能与肝癌的形成有关。
Objective To study the regulatory effect of Hepatitis C Virus F protein on Bcl-2 and Bax.Methods Full length F gene of HCV was amplified by PCR,using the plasmid containing HCV 1b Full length open reading frame as template.HCV F gene-containing,plasmid pEGFP-C3-F and HCV core gene-containing pEGFP-C3-C were constructed and transiently transfected into HepG2 cells.48h posttransfection,Western blotting was used to determine the changes at translation levels of Bcl-2 and Bax genes.Results The levels of Bcl-2 expression were a little higher in pEGFP-C3-F transfected HepG2 cells than that of non-transfected cells,but significantly higher levels of Bax expression.Conclusions F protein is of regulatory properties in celluar oncogen Bcl-2 and anti-oncogen Bax,which may be implicated in the formation of hepatocelluar carcinoma.