腺病毒为载体的HSV1-TK基因对肿瘤细胞系抑制作用的研究

Study on Killing Effect of Tumor in Vitro with Adenovirus Vector Containing HSVI-TK Gene

目的:用E1区缺失的含HSV1-TK基因和CMV启动子的重组腺病毒AdE1CMVHSV1-TK(AdTK)前体药物GCV即AdTK/GCV系统,观察其在体外对PG49、H460、MCF-7、Hela4种建株肿瘤细胞系的抑制作用、旁杀伤效应和AdTK/GCV系统处理的肿瘤细胞培养上清液对肿瘤细胞的杀伤效应。方法;采用MTT法测定细胞生长抑制率,AdLacZ作为对照病毒。结果:细胞生长抑制率随重组病毒的浓度、前体药物的浓度及作用时间的增加而升高,而对照病毒则没有显示出对肿瘤细胞明显的抑制作用。旁杀伤效应显示,细胞抑制率随导入AdTK病毒的肿瘤细胞比例的增加而升高。另外肿瘤细胞呈现出上清液浓度依赖性抑制,其生长抑制率随上清液浓度的增加而升高。结论:AdTK/GCV系统对4种肿瘤细胞系有明显的抑制作用及旁杀伤效应。

Objective: Cytotoxic effects on PG49, H460, MCF-7, Hela established tumor cells with recombinant replication-defective adenovirus (AdE, CMVHSV1-TK) containing HSV1-TK gene and CMV promoter with EI region delete i. e. AdTK/GCV system were assayed. The bystander effect was measured as well as the killing effect of the reactive supernatant in tumor cells treated with AdTK/GCV system. Methods: Using MTT method with AdLacZ being control. Results: The growth inhibition rate depended on AdE, CMVTK dose, the concentration of GCV, and the time of interaction. There was no significant effect in TK-negative tumor cells. Considerable bystander effect was observed. The killing effect resulted from the two methods revealed that the growth inhibition rate increased with the raising of the ratio of TK-positive cells as well as the concentration of the supernatant from tumor cells treated with AdTK/GCV system. Conclusion: AdTK/GCV system has a considerable inhibit and bystander effect on tumor cells.