端粒酶RNA特异性核酶抑制卵巢癌细胞端粒酶的活性

Attenuation of telomerase activity by a hammerhead ribozyme targeting the template region of telomerase RNA in ovarian carcinoma cell

目的 研究端粒酶 RNA特异性核酶对卵巢癌细胞活性的抑制作用 .方法 用脂质体法将构建好的核酶逆转录病毒载体转染至卵巢癌 HO- 8910细胞 ,以 MTT比色法检测细胞的生长 ,应用流式细胞仪分析细胞周期细胞 ,观察细胞的增殖情况 ,同时扩增端粒重复序列 ,结合杂交分析 ,检测细胞的端粒酶活性 .结果 MTT检测发现转染核酶后的细胞生长的潜伏期延长 (48→ 72 h) ,对数生长期减缓 (4→ 5 d) ,流式细胞仪结果显示其 G1期比例明显增高 (70 .0 %→ 80 .5 % ,P<0 .0 1) ,S期细胞比例明显降低 (19. 0 %→ 13. 7% ,P<0 .0 1) ,细胞增殖下降 ,细胞的端粒酶活性亦降低 .结论 端粒酶 RNA特异性核酶能抑制卵巢癌细胞端粒酶活性 ,抑制细胞增殖 .

AIM To explore the effects of hammerhead ribozyme against human telomerase RNA in ovarian carcinoma cells. METHODS We transfected the ribozyme directed against the RNA component of human telomerase into HO 8910, human ovarian carcinoma cells. Then we observed the propagation of the cells with MTT, analyzed cell cycle phases under FCM and detected their telomerase activity by TRAP Hyb Kit. RESULTS After transfection, we found that the latent period of the transfected cells prolonged (48→72 h) and the logarithm growth period also lengthened (4→5 d). FCM showed that cells in G1 phase increased obviously (70.0% →80.5%, P <0.01) and decreased obviously in S phase (19.0%→13.7%, P <0.01). The propagation of the cells was inhibited and the telomerase activity was reduced. CONCLUSION Telomerase ribozyme inhibites the telomerase activity in ovarian carcinoma cells.