两个不同筛选特性的表达HBV X基因肝癌细胞模型的建立

目的：构建两个不同筛选特性的表达HBV X基因肝癌细胞模型。方法：应用脂质体介导转染克隆有HBV X全基因真核表达载体pCEP4-X和pcDNA3.1(+)-X入肝癌细胞HepG2，hygromycin和neomycin筛选单细胞克隆，传代培养一定时期后应用PCR,RT-PCR和Western blot方法鉴定HBV X基因的表达。结果：转染后成功筛选出耐hygromycin或neomycin的单细胞克隆，并能传代培养一定时期。PCR,RT-PCR和Western blot结果显示HBV X基因获得表达。结论:成功构建不同筛选特性的两个HBV X基因表达肝癌细胞模型。

Construction of two hepatocellular carcinoma cell models for the expression of HBV X gene with different selection characteristics

Objective To construct 2 hepatocellular carcinoma(HCC) cell models for the expre-(ssion) of HBV X gene with different selection characteristics.Methods HepG2 HCC cells were infected with eukaryotic expression vectors with HBV X gene,pCEP4-X,and pcDNA3.1( )-X.Single cell clone was selected by hygromycin and neomycin.After propagating culture for certain periods,the HBV X gene expression was identified by PCR,RT-PCR,and Western blot.Results Single HCC cell clone with HBV X gene transferred resistant to hygromycin and neomycin was selectively cultured,and the cells could be propagated for certain periods.PCR,RT-PCR,and Western blot identified the expression of HBV X gene.Conclusion Two HCC cell models for the expression of HBV X gene with different selection characteristics have been successfully constructed.