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目的 探讨HSV TK/GCV自杀基因系统对小鼠乳腺癌细胞系MA782 / 5S 810 2体外及体内杀伤作用及其产生的旁观者效应。方法 采用脂质体转染法将GINaTK载体转入包装细胞PA317。取病毒上清液感染小鼠乳腺癌细胞MA782 / 5S 810 2 ,得到带有HSV TK基因的MA782 / 5S 810 2 /TK细胞 ,并将其分别用于体外和体内实验。结果 载体HSV TK导入了PA317细胞。体外实验结果显示 ,当MA782 / 5S 810 2 /TK细胞数占混合细胞 10 %时 ,低浓度 (10 μg/ml)的GCV就可将 5 0 %左右的肿瘤细胞杀死。体内实验结果显示GCV可明显抑制MA782 / 5S 810 2 /TK细胞在BALB/C小鼠体内的肿瘤形成。实验组肿瘤组织与对照组相比存在明显的病理学改变。结论 逆转录病毒可介导HSV TK基因转入小鼠乳腺癌细胞MA782 / 5S 810 2并获稳定表达 ,HSV TK/GCV自杀基因系统在体内外对乳腺癌细胞均有杀伤作用 ,且存在明显的旁观者效应。
Objective To investigate the in vitro and in vivo killing effects of HSV TK / GCV suicide gene system on murine breast cancer cell line MA782 / 5S 810 2 and its bystander effect. Methods The GINaTK vector was transfected into packaging cell PA317 by lipofection. The virus supernatant was used to infect mouse breast cancer cells MA782 / 5S 810 2 to obtain MA782 / 5S 810 2 / TK cells bearing the HSV TK gene and used for in vitro and in vivo experiments, respectively. Results Vector HSV TK was introduced into PA317 cells. In vitro experiments showed that about 50% of the tumor cells could be killed by GCV at a low concentration (10 μg / ml) when MA782 / 5S 810 2 / TK cells accounted for 10% of the mixed cells. In vivo experimental results show that GCV can significantly inhibit the tumor formation of MA782 / 5S 810 2 / TK cells in BALB / C mice. There were obvious pathological changes in the experimental group compared with the control group. Conclusion The retrovirus can transfer HSV TK gene into mouse mammary carcinoma cell line MA782 / 5S 810 2 and get stable expression. The HSV TK / GCV suicide gene system has killing effect on breast cancer cells both in vitro and in vivo, Bystander effect.