沉默HeLa细胞端粒酶活性对其在裸鼠体内移植瘤生长的影响  

Effects on xenografts in nude mice of HeLa cells after silencing telomerase activity by RNA interference

作　　者：石英爱[1] 赵强[2] 张海英[1] 张丽红[1] 王光兰[1] 陈爽[1] 王勇[3] 

机构地区：[1]吉林大学基础医学院病理生物学教育部重点研究室,吉林长春130021 [2]吉林大学第一临床医学院儿外科 [3]吉林大学第一临床医学院心外科

出　　处：《中国实验诊断学》2009年第9期1152-1154,共3页Chinese Journal of Laboratory Diagnosis

基　　金：教育部高校优秀青年教师奖励计划(2000134);吉林省卫生厅重点实验室科研项目;吉林省科技发展计划项目资助课题(200705369)

摘　　要：目的沉默HeLa细胞人端粒酶催化亚单位(human telomerase catalytic subunit,hTERT)基因、抑制细胞端粒酶活性,观察细胞生长状态和其对裸鼠体内移植接种成瘤能力的影响。方法RNAi技术,脂质体转染,筛选沉默hTERT基因稳定转染细胞株和无干扰效果的对照细胞株,端粒重复序列扩增-酶联免疫吸附测定法(TRAP-ELISA)测定所筛选的细胞株端粒酶活性;进一步绘制各细胞株的生长曲线;裸鼠移植接种实验观察各株细胞在裸鼠体内成瘤能力。结果成功获得稳定转染具有沉默hTERT基因,端粒酶活性明显降低的实验组细胞株2个即T1,T2,同时获得对照细胞株1个(N1),还以无转染的HeLa细胞为亲本对照。细胞生长曲线显示与相应的对照组相比,实验组细胞(T1,T2)生长速度明显减慢(P<0.05)。裸鼠体内成瘤实验结果显示实验组和对照组均有皮下肿瘤形成,移植接种20天后,处死裸鼠取出肿瘤称量结果显示对照组形成的实体肿瘤平均重量分别为:0.057 7±0.034 9 g和0.054 4±0.033 5 g,而实验组T1和T2组肿瘤重量明显减轻(P<0.05),仅分别为:0.024 5±0.010 9 g和0.027 2±0.007 5 g。结论沉默Hela细胞hTERT基因降低肿瘤细胞的端粒酶活性,使细胞的体外增殖速度减慢,在裸鼠体内皮下成瘤能力受到明显抑制。Objective To observe the effects on the growth of HeLa cells and xenografts in nude mice after silencing human telomerase catalytic subunit （hTERT） gene and telomerase activity by RNA interference. Methods RNA interference and lipid formulation were utilized to screen the stable cell lines with telomerase activity inhibition. The telomerase activity of cell hnes was evaluated by telomeric repeat amplification protocol （TRAP） ELISA （TRAP-ELISA）. Cell proliferation was investigated by cell growth curve. Tumor growth in vivo was estimated by the volume and weight of xenografts in nude mice. Results Two stable cell lines with silenced hTERT gene and lowered telomerase activity （T1 ,T2） were chosen as experimental groups. One sable cell line with normal hTERT gene （N1）,and normal HeLa cells were chosen as control groups. The cell proliferation ability of experimental groups was decreased compare with control groups （ P 〈 0.05 ）. There were tumor forrmation in both of experimental and control groups. After 20 days, all the nude mice were sacrificed. The weights of xenugrafts in nude mice were 0. 057 7 ± 0. 034 9 g and 0. 054 4 ± 0. 033 5 g in experimental groups （T1 ,T2） ,and 0.024 5±0.010 9 g and 0.027 2 ± 0.0375 g in control groups respectively.There was significant difference between them （ P 〈 0.05）. Conclusion After silencing the hTERT gene of HeLa cells, the activity of telomerase can be inhibited, and the cell proliferation showed predominantly reduced, and also the weights of xenografts in nude mice diminished obviously.

关 键 词：端粒酶 HTERT RNA干涉 裸鼠 

分 类 号：R73-3[医药卫生—肿瘤]