A549肿瘤细胞中特异性hTERT-siRNA的干扰作用研究  

RNA interference directed by small interference RNA of hTERT expressed in lung carcinoma cell A549

作　　者：赵亚力[1] 韩卫东[1] 宋海静[1] 李琦[1] 马学斌[1] 郝好杰[1] 

机构地区：[1]解放军总医院,北京100853

出　　处：《解放军医学杂志》2004年第8期670-673,共4页Medical Journal of Chinese People's Liberation Army

基　　金：北京市自然科学基金 (编号 70 4 2 0 59)资助课题

摘　　要：目的　研究hTERT特异的siRNA对肿瘤细胞端粒酶活性及细胞生长的抑制作用。方法　设计和构建由U6启动子驱动的产生特异hTERT的siRNA表达质粒 ,转染至A5 4 9肺肿瘤细胞 ,以TRAP ELISA方法观察细胞端粒酶活性 ,Westernblot检测端粒酶蛋白的表达 ,MTT法检测对细胞生长的影响。结果　构建的针对hTERT基因 74 5靶位点的U6表达质粒具有明显的干扰效果 ,受到特异hTERT siRNA干扰的A5 4 9肿瘤细胞端粒酶表达下调 ,细胞生长受抑。结论　应用RNA干扰技术可阻止hTERT基因的表达 ,有望成为肿瘤基因治疗新的研究方向。Objective To investigate the inhibitory effect of siRNA human telomerase transcriptase (hTERT) on activity of telomerase and proliferation of lung carcinoma cell A549. Methods A plasmid including U6 promoter and siRNA of hTERT was designed and constructed. The plasmid was transfected into A549 cell line. The telomerase activity was tested by telomerase repeat amplification protocol ELISA (TRAP-ELISA). MTT assay was used to assay the cell proliferation activity,and hTERT expression was assessed by Western blot. Result The U6 expression plasmid that was constructed for hTERT gene 745 showed obvious interfering effect. hTERT-siRNA could down-regulate the expression of hTERT protein,inhibit telomerase activity and proliferation of A549 cells. Conclusion siRNA of hTERT can inhibit the expression of human telomerase and proliferation of A549 cells. It may open a new approach to the use of siRNA as a new tool to study gene function in cancer cell lines,and may be developed to be a new gene therapeutic agent for cancer.

关 键 词：肿瘤细胞 端粒酶 RNA干扰 

分 类 号：R730.5[医药卫生—肿瘤]