丙型肝炎病毒核心蛋白在HepG2细胞中的表达及其对端粒酶活性的影响

HCV core protein expression in HepG2 cells and its effect on telomerase activity

目的 建立HCV核心蛋白细胞表达模型 ,并探讨其对细胞端粒酶活性的影响。方法用PCR法扩增出HCV核心基因cDNA ,将其插入真核表达载体pBK CMV的HindⅢ和BamHⅠ位点间 ,构建重组质粒pBK HCVc。再将重组质粒pBK HCVc和空载体分别导入肝癌细胞株HepG2中 ,G418筛选 ,RT PCR、免疫组化和蛋白印迹鉴定HCV核心蛋白表达。PCR ELISA法检测端粒酶活性。结果 构建的pBK HCVc质粒在HepG2细胞中有稳定表达。表达HCV核心蛋白的细胞HepG2 C的端粒酶活性较转染空载体的细胞HepG2 CMV明显升高。结论 HCV核心蛋白上调了端粒酶活性 ,可能是HCV诱发肝细胞癌的一种途径。

Objective To establish an experimental model of HCV core protein expression and explore its effect on telomerase activity. Methods The HCV core gene cDNA was recoverd by PCR, and cloned into pBK-CMV. The recombinant plasmid(pBK-HCVc) and the vector-alone were transfected into HepG2 cells with liposome. After been selected with G418, resistant colonies were obtained. The reverse transcription PCR, inmmunocytochemistry and Western blot were used to analyze HCV core protein expression. Telomerase activity was analyzed by PCR-ELISA. Results The recombinant plasmid express HCV core protein efficiently under the control of vector′s promoter. The telomerase activity of HepG2-C cells is higher than HepG2-CMV. Conclusion HCV C protein increases the telomerase activity of HepG2 cells. It is suggested that HCV C protein contribute to viral carcinogenesis.