两个不同筛选特性的表达HBV X基因肝癌细胞模型的建立

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

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,hygromyc in和neomyc in筛选单细胞克隆,传代培养一定时期后应用PCR,RT-PCR和W estern b lot方法鉴定HBV X基因的表达。结果:转染后成功筛选出耐hygromyc in或neomyc in的单细胞克隆,并能传代培养一定时期。PCR,RT-PCR和W estern b lot结果显示HBVX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。

Objective To construct 2 hepatocellular carcinoma （HCC） cell models for the expression 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.