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目的观察双自杀基因胞嘧啶脱氨酶基因(CD)和单纯疱疹病毒胸苷激酶基因(HSV-tk)对胆管癌细胞的抑制作用以及转染缝隙连接蛋白43(Cx43)基因对旁观者效应的影响。方法四甲基偶氮唑蓝(MTT)法测定双自杀基因系统作用下和/或Cx43基因转染后胆管癌QBC939细胞的细胞抑制率及旁观者效应。RT-PCR检测双自杀基因和Cx43基因的表达。Westernblot检测Cx43蛋白的表达。结果CD和tk基因在QBC939/CD+tk细胞和QBC939/CD+tk+Cx43细胞中获得稳定表达。mRNA和蛋白水平均显示转染Cx43基因的QBC939/CD+tk+Cx43细胞的Cx43的表达量明显增加。双自杀基因系统对QBC939/CD+tk+Cx43细胞组的抑制作用较QBC939/CD+tk细胞组高。结论双自杀基因系统对胆管癌细胞有明显的抑制作用,Cx43基因的导入可明显增强旁观者效应,提高对肿瘤细胞的抑制作用。
Objective To observe the inhibitory effect of double suicide gene cytosine deaminase (CD) gene and herpes simplex virus thymidine kinase gene (HSV-tk) on cholangiocarcinoma cells and the effects of transfection of connexin43 (Cx43) gene on bystander effect influences. Methods MTT assay was used to determine the cell inhibitory rate and bystander effect of double suicide gene system and / or Cx43 gene transfection in cholangiocarcinoma QBC939 cells. RT-PCR double Suicide gene and Cx43 gene expression. Western blot was used to detect the expression of Cx43 protein. Results The CD and tk genes were stably expressed in QBC939 / CD + tk cells and QBC939 / CD + tk + Cx43 cells. The mRNA and protein levels of Cx43 in QBC939 / CD + tk + Cx43 cells transfected with Cx43 gene were significantly increased. Double suicide gene system on QBC939 / CD + tk + Cx43 cell group than QBC939 / CD + tk cell group. Conclusion Double suicide gene system has significant inhibitory effect on cholangiocarcinoma cells. The introduction of Cx43 gene enhances the bystander effect and enhances the inhibitory effect on tumor cells.