Wu Wen-bin, Shao Sheng-wen, Zhao Lan-juan, Luan Jie, Cao Jie, Gao Jun, Zhu Shi-ying, Qi Zhong-tian
Department of Microbiology, State Key Laboratory of Medical Immunology, Second Military Medical University, Shanghai, PR China.
Intervirology. 2007;50(5):341-6. doi: 10.1159/000107271. Epub 2007 Aug 15.
Hepatitis C virus (HCV) F protein is a newly identified protein encoded by an alternative open reading frame that +1 overlaps core-encoding gene. It has been found that regulation of c-myc and p53 genes by HCV core protein is involved in liver cancer genesis. We wondered whether HCV F protein exerts similar or adverse regulatory effects on the transcription of c-myc and p53 genes.
HCV F gene-containing, plasmid pcDNA3.1-F and HCV core gene-containing pcDNA3.1-C were constructed and transiently transfected into HepG(2) cells. Real-time quantitative PCR or Western blotting was used to determine the changes at transcription or translation levels of c-myc and p53 genes.
The transcription level of c-myc was much higher in pcDNA3.1-F transfected cells than those without plasmid transfected. Whereas the level of p53 transcription in pcDNA3.1-F transfected cells was lower than those in the parental cells. Moreover, levels of c-myc expression were up-regulated and those of p53 expression were down-regulated by HCV F protein.
HCV F protein is of regulatory properties in cellular oncogene c-myc and anti-oncogene p53, which may be implicated in the formation of hepatocellular carcinoma.
丙型肝炎病毒(HCV)F蛋白是一种新发现的由替代开放阅读框编码的蛋白,其+1重叠于核心编码基因。已发现HCV核心蛋白对c-myc和p53基因的调控参与肝癌发生。我们想知道HCV F蛋白对c-myc和p53基因转录是否发挥类似或相反的调控作用。
构建含HCV F基因的质粒pcDNA3.1-F和含HCV核心基因的pcDNA3.1-C,并将其瞬时转染至HepG(2)细胞。采用实时定量PCR或蛋白质印迹法检测c-myc和p53基因转录或翻译水平的变化。
pcDNA3.1-F转染细胞中c-myc的转录水平远高于未转染质粒的细胞。而pcDNA3.1-F转染细胞中p53的转录水平低于亲本细胞。此外,HCV F蛋白上调c-myc表达水平,下调p53表达水平。
HCV F蛋白对细胞癌基因c-myc和抑癌基因p53具有调控作用,这可能与肝细胞癌的形成有关。