PBI-SUR-TK载体靶向介导HSV-TK自杀基因诱导肝癌细胞凋亡

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HepG₂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。

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

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 transfect 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.