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目的 :观察单纯疱疹病毒胸苷激酶 (HSV TK)丙氧鸟苷 (GCV)自杀基因系统对人宫颈癌Hela细胞系体外杀伤作用及其旁观效应。 方法 :采用脂质体转染法将GINaTK载体转入包装细胞PA317。取病毒上清液感染Hela细胞 ,得到带有HSV TK基因的Hela/TK细胞 ,并将其用于体外实验。 结果 :载体HSV TK导入了PA317细胞。体外实验结果显示 ,当Hela/TK细胞数占混合细胞 10 %时 ,低浓度 (10mg/L)的GCV就可将 5 0 %左右的肿瘤细胞杀死。GCV作用后的实验组Hela /TK细胞基因组经琼脂糖凝胶电泳后可观察到典型的DNA梯状条带。 结论 :逆转录病毒可介导HSV TK基因转入人宫颈癌Hela细胞并获稳定表达 ,HSV TK/GCV自杀基因系统在体外对宫颈癌细胞有杀伤作用 ,且存在明显的旁观效应。
OBJECTIVE: To observe the killing effect of HSV TK gonadotrophin (GCV) suicide gene system on human cervical carcinoma Hela cell line and its bystander effect. Methods: GINaTK vector was transfected into packaging cell PA317 by lipofection method. Hela cells were infected with the virus supernatant to obtain Hela / TK cells with the HSV TK gene and used for in vitro experiments. Results: The vector HSV TK was introduced into PA317 cells. In vitro experiments showed that when Hela / TK cells accounted for 10% of mixed cells, low concentration (10mg / L) of GCV can kill about 50% of tumor cells. GCV-treated experimental group Hela / TK cell genome after agarose gel electrophoresis typical DNA ladder can be observed. CONCLUSION: The retrovirus can transfer HSV TK gene into human cervical cancer Hela cells and get stable expression. HSV TK / GCV suicide gene system can kill cervical cancer cells in vitro, and there is obvious side effect.