pcDNA3.1(+)/hTERT-tk真核表达质粒的构建及其在HepG-2细胞中的表达

Construction of pcDNA3.1(+)/hTERT-tk Eukaryotic Expression Plasmid and Its Expression in HepG-2 Cells

目的:研究人端粒酶逆转录酶(hTERT)基因核心启动子调控的人单纯疱疹病毒胸苷激酶基因/更昔洛韦(HSV-tk/GCV)系统对肝癌HepG-2细胞的体外杀伤作用。方法:构建hTERT基因启动子调控的tk基因真核表达载体pcDNA3.1(+)/hTERT-tk,用脂质体法分别转染肝癌细胞(HepG-2)和正常肝细胞(L-02)后给予更昔洛韦(GCV),用TUNEL法观察自杀基因对肝癌细胞生长的影响。结果:HETRT启动子调控下的HSV-tk/GCV系统对肝癌HepG-2细胞有明显的杀伤作用,而对正常肝细胞则作用不明显。结论:hTERT-tk/GCV基因体系能够靶向杀伤肝癌细胞,有靶向治疗肝癌的潜力。

Objective：To observe the killing effect of HSV-tk/GCV system driven by human telomerase reverse transeriptase（hTERT）core promoter against human hepatocellular carcinoma cell line HepG-2 in vitro. Methods：pcDNA3.1 （＋）/ hTERT-tk eukaryotic expression plasmid containing HSV-tk gene under the control of hTERT core promoter was constructed and was transfected into hepatoma cells （HepG-2）and normal hepatic cells （L-02）by liposome method. Then transfected cells were selected with G418, at last GCV was added and cell growth was observed by Tunnel. Results： HSV-tk/GCV system could significantly inhibited the growth of HepG-2 cell, and had little cytotoxieity to normal hepatic cell. Conclusion ： HSV-tk/GCV system controlled by hTERT core promoter has selectively killing effect on HepG-2 cell in vitro.