摘要
目的观察胞嘧啶脱氨酶(CD)基因和单纯疱疹病毒胸苷激酶(HSV-tk)基因对胆管癌细胞和胆管癌移植瘤的作用以及转染间隙连接蛋白43(Cx43)基因对旁观者效应的影响。方法MTT法比较双自杀基因系统作用下和(或)Cx43基因转染后QBC939细胞的细胞抑制率及旁观者效应。前体药物应用前、后测量移植瘤的大小,判断自杀基因系统的作用和Cx43蛋白对旁观者效应的影响。结果CD和HSV-tk基因在转染细胞中获稳定表达。mRNA和蛋白水平均显示CD+tk+Cx+细胞中Cx43的表达量增加。自杀基因系统对CD+tk+Cx+细胞组和它生成的移植瘤组的抑制作用均比CD+tk+细胞组高。结论双自杀基因系统在体内、外对胆管癌细胞和移植瘤均有明显的抑制作用,Cx43基因导入可明显增强旁观者效应和对肿瘤的抑制作用。
Objective To observe the influence of connexin 43 （Cx43） on bystander effects induced by cyto sine deaminase （CD） and herpes simplex virus thymidine kinase （HSV-tk） coexpression suicide genes system in hu-man cholangiocarcinoma QBC939 cells and transplantation tumors in nude mice. Methods In vitro, the CD^＋ tk^＋ and CD^＋tk^＋Cx^- cells were respectively treated with 5-fluorocytosine （S-FC） and ganciclovir （GCV）. The cytotoxic efficacy was evaluated by microculture tetrajolium test （MTT） method. In order to investigate the influence of Cx43 on bystander effects, the volumes of transplantation tumors of the CD^＋ tk^＋ and CD^＋ tk^-Cx^- cells were measured before and after application of 5-FC and GCV. Results CD and tk gene were stably expressed in transfected QBC939 cells. Increasing expression of Cx43 was determined by testing for the presence of Cx43 mRNA by RT-PCR and the presence of Cx43 protein by Western blot in CD^-tk^-Cx^＋ cells. The killing effect of S FC and GCV on CD^-tk^＋ Cx^＋ cells was more effective than that on CD^＋ tk^＋ ceils both in vitro and in vivo. Conclusion Double suicide genes system CD/5-FC＋tk/GCV could induce remarkable killing effect on cholangiocarcinoma cells in vitro and transplantation tumors in vivo. The cotransfection of Cx43 gene is able to enhance the bystander effects and the inhibition of carcinoma cells.
出处
《中国普外基础与临床杂志》
CAS
2008年第8期583-588,共6页
Chinese Journal of Bases and Clinics In General Surgery
基金
国家863高技术研究发展计划资助项目(编号:2002AA214061)~~
