摘要
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 investigate the repressing effect on human cholangioearcinoma QBC939 cells with cytosine deaminase（CD） and herpes simplex virus thymidine kinase（HSV- tk） double suicide genes system and the influence of the transduction of connexin 43 to bystander effect. Methods QBC939/CD ＋ tk cells and QBC939/CD ＋ tk ＋ Cx43 cells were respectively treated with 5 - fluorocytosine （5 - Fc） and Ganciclovir（GCV）, the cytoxicity efficacy was evaluated by microculture tetrajolium test （MTT） method. The expression of CD, HSV- tk and Cx43 gene were detected by RT- PCR. The expression of Cx43 protein was detected by Western blot. Results Double suicide genes were stably expressed in QBC939/CD ＋ tk cells and QBC939/CD ＋ tk ＋ Cx43eells. At mRNA and protein level, the degree of expression of Cx43 in QBC939/CD ＋ tk ＋ Cx43 cells were increased after transduction of Cx43 gene. The repressing capability of 5 - Fc and GCV on QBC939/CD ＋ tk ＋ Cx43 cells was more effective than that on QBC939/CD ＋ tk cells. Conclusion The repressing effect of double suicide genes system to cholangiocarcinoma cells is strong and the bystander effect is observed. The repressing effect is improved and bystander effect is stronger after the transduction of Cx43 gene.
出处
《临床外科杂志》
2006年第3期154-156,共3页
Journal of Clinical Surgery
基金
国家863高技术研究发展计划资助项目(编号:2002AA214061)
