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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 an expression vector regulated by Survivin promoter and to detect the effect of HSV-TK suicide gene on the apoptosis of hepatocellular carcinoma cell line HepG_2 and normal liver cell line HL-7702 under the control of promoter. Methods: Plasmid PBI-SUR-TK containing TK gene was synthesized and transfected into hepatoma cells and hepatocytes by Lipofectamine 2000. The results showed that PBI- Then, the expression of genes and proteins were detected by RT-PCR and Western blot respectively. The proliferation of cells was detected by CCK8 assay and the apoptosis was detected by flow cytometry. Results: There were more TK gene expression products in the transfection group of liver cancer cells, the proliferation was weakened and the apoptosis was obvious. Conclusion: Survivin promoter-driven HSV-TK / GCV suicide gene system may have a therapeutic effect on hepatocellular carcinoma.