丙型肝炎双移位F蛋白抑制肝癌细胞p16、p21的表达

HCV DF protein inhibits expression of p16 and p21 in HepG_2 cells

目的:探讨丙型肝炎双移位F(DF)蛋白对肝癌细胞抑癌基因p16、p21转录与表达的影响。方法:PCR扩增HCV1b型DF基因,构建pCDNA3.0/HCV-DF真核表达载体。再转染至肝癌细胞HepG2中,G418筛选稳定表达细胞株,Western blot检测p16、p21蛋白表达及半定量RT-PCR法检测p16、p21基因转录,并以pCDNA3.0空质粒作为阴性对照。结果:重组质粒pCDNA3.0/HCV-DF蛋白在HepG2细胞中稳定表达,pCDNA3.0/HCV-DF转染细胞中p16、p21 mR-NA转录水平和蛋白表达水平较空质粒转染的细胞明显下降。结论:HCV-DF蛋白能够抑制p16、p21表达,提示可能参与肝细胞癌变发生发展。

AIM： To investigate the effect of HCV DF（Double-shift F） protein on the expression p16 and p21 in HepG2 cells.METHODS： DF gene was amplificated from the whole HCV 1b genome,and cloned into pCDNA3.0 vecter.The recombinant plasmid（pCDNA3.0/HCV-DF） and empty vector were transfected into HepG2 cells.Screening was performed with G418.p16 and p21 mRNA were detected by semi-quantitative RT-PCR,and protein by Western blot.RESULTS： Stable expression of the recombinant plasmid was found in HCV DF protein.The expression of p16 and p21 in HepG2 cells transfected with pCDNA3.0/HCV-DF were lower than those with blank plasmid.CONCLISION： HCV DF protein inhibits expression of p16 and p21 in HepG2 cells.This suggested that HCV DF protein may participate in the progress of hepatocellular carcinoma.