RNAi沉默人端粒酶逆转录酶对HeLa细胞生长及药物敏感性影响的研究.pdfVIP

中华肿瘤防治杂志 2009 年 6 月第 16 卷第 11 期 　　CH IN J CAN CER PR EV TR EA T ,J une 2009 ,Vol 16 　No 11 ·829 · 实验研究 RNAi 沉默人端粒酶逆转录酶对 HeL a 细胞生长 及药物敏感性影响的研究 米洋 , 　廖 　晶 , 　郭雪玲 , 　黄喜阳 , 　黄小荣 , 　梁昌盛 , 　周俊宜 中山大学中山医学院生化教研室 ,广东 广州 5 10080 Effect of silencing hTERT by RNAi on growth and drug sen sitivity of HeLa cell s M I Yang , L IA O M inj ing , GUO X ueling , H UA N G X iy ang , H UA N G X iaorong , L IA N G Changs heng , Z H OU J uny i D ep a rtment of B iochem is t ry , Z hongs han M e d ical Col leg e , S un Yatsen Uni vers i ty , Guang z hou 510080 , P. R . Chi na 【摘要】　目的:探讨利用短发夹 RN A ( short [ ABSTRACT] 　OBJECTIVE : To explore t he effect of silencing hairpin RNA , shRN A) 抑制人端粒酶逆转录 short hairpin RN A ( sh RN A) t ar geting h T ER T on HeL a cell s. ( ) 酶 h T ER T 的表达后对人宫颈癌 HeL a 细胞 METHODS : Exp ression vector s generating shRN A t ar geting 生长及药物敏感性的影响 。方法 : 将构建的 h T ER T were con st ruct ed and int ro duced into HeL a cell s by lipo h T ER Tsh RN A 真核表达载体用脂质体转染 some . St able cell st rain s were gained by G4 18 screening and 入 HeL a 细胞中 ,经 G4 18 筛选得到稳定转染 t heir growt h velocities were mea sured . The exp ression of 的细胞株 ,并绘制细胞生长曲线 。R TPCR 和 h T ER T and cmyc were analyzed by R TPCR and West ern blot . 蛋白质印迹法分别检测 h T ER T 、cmyc mR The respon se of h T ER Tsilencing cell s and t he cont rol group to N A 和蛋 白表达 。镜下观察干扰组细胞与对 chemot herap eutic drugs was ob served . Flow cytomet ry and im 照组细胞对化疗药物的反应 ,并在紫杉醇给药 munochemi st ry were u sed to analyze t