PBI-SUR-TK载体靶向介导HSV-TK自杀基因诱导肝癌细胞凋亡

Targeted Treatment of Hepatoma Using HSV-TK Suicide Gene with The PBI-SUR-TK Vector

目的:构建Survivin启动子调控的表达载体,并检测在启动子调控下HSV-TK自杀基因对肝癌细胞HepG_2和正常肝细胞HL-7702凋亡的影响。方法:合成含TK基因的质粒PBI-SUR-TK,利用脂质体Lipofectamine 2000将其导入肝癌细胞和肝细胞。然后分别运用RT-PCR和Western blot特异性检测基因和蛋白的表达情况;利用CCK8方法检测细胞增殖情况,流式细胞仪上机检测细胞凋亡情况。结果:肝癌细胞转染组有更多的TK基因表达产物,增殖情况减弱,凋亡情况明显。结论:Survivin启动子驱动的HSV-TK/GCV自杀基因系统对肝癌可能有一定的治疗作用。

Objective： To construct a kind of expression vector which is cotrolled by survivin promoter, making a survivin promotor driving HSV-TK/GCV suicide gene system, and then detect its apoptosis inducing effect on HepG2 and HL-7702 cells. Methods ： The study group synthesized a plasmid PBI-SUR-TK which contains TK gene, and transfeet it into HepG2 and HL-7702 cells by liposome Lipofectanfine 2000. The HSV-TK gene expression by RT-PCR and the proteinlevel by Western blotting were detected. After adding GCV, CCK8 and flow cytometry were used to analyse the cell proliferation and apoptosis. Results： Contrasted with the control group, the HepG2 experimental group expressed more TK gene products,less proliferation and more apoptosis. There was no obvious difference between the HL-7702 groups. Conclusion： The HSV-TK/GCV suicide gene system driven by survivin promoter possibly has therapeutical effects on hepatoma.