核酶对食管癌EC9706细胞端粒酶活性及凋亡的影响

Inhibitory effects of ribozyme on telomerase activity and apoptosis of esophageal carcinoma cell EC9706

目的 :观察针对人端粒酶RNA模板区的核酶对食管癌细胞端粒酶活性和细胞凋亡的影响。方法 :利用脂质体Lipofectamine介导 ,将已构建好的带有端粒酶核酶基因的重组质粒pBBS2 12Rz及空载质粒pBBS2 12转染食管癌EC970 6细胞 ,采用TRAP ELISA法检测端粒酶活性 ,用倒置相差显微镜及流式细胞仪观察细胞生长和凋亡情况。结果 :重组质粒pBBS2 12Rz转染的食管癌EC970 6细胞的端粒酶活性明显下降 ,细胞生长速度明显变慢 ,凋亡加速。结论 :端粒酶核酶对食管癌细胞端粒酶活性和细胞生长有抑制作用 。

OBJECTIVE:To observe the effects of the ribozyme targeting the human telomerase RNA (hTR) on telomerase activity and cell apoptosis of esophageal carcinoma cell line EC9706.METHODS:The eucaryotic expression vector pBBS212Rz and vacant plasmid pBBS212 as a control were transfected respectively into the human esophageal carcinoma cell line EC9706 by lipofectamine.Telomerase activity of transfected cells was detected by TRAP-ELISA (Telomeric Repeat Amplification Protocol and enzyme-linked immunosorbant assay).Cell proliferation and apoptosis were detected by phase contrast microscope and flow cytometry.RESULTS:Telomerase activity of ribozyme transfected group decreased obviously compared with the control group.Cell proliferation was became slow obviously and cell apoptosis was accelerated.CONCLUSIONS:Those findings suggested that the hammerhead ribozyme targeting the hTR template can inhibit telomerase activity of esophageal carcinoma in vitro and it will be a new therapeutic approach to esophageal carcinoma.