RNA干扰对膀胱移行细胞癌BIU-87细胞株端粒酶活性及其增殖的影响  

Effects of siRNA on Telomerase Activity and the Growth Inhibition in Transitional Cell Carcinoma of Bladder BIU-87 Cell Line in Vitro

作　　者：谢海龙[1] 谭桂煌[1] 刘重元[1] 程文[2] 

机构地区：[1]南华大学肿瘤研究所,湖南衡阳421001 [2]南京军区南京总医院泌尿外科

出　　处：《南华大学学报（医学版）》2009年第2期161-165,共5页Journal of Nanhua University(Medical Edition)

基　　金：国家自然科学基金(30572030)

摘　　要：目的探讨针对人端粒酶RNA(hTR)及其催化亚基(hTERT)的小干扰RNA(siRNA)对膀胱移行细胞癌细胞株BIU-87端粒酶活性及其增殖的影响。方法根据人TR mRNA和TERT mRNA的序列,分别设计合成3对不同的且能干扰TR和TERT基因表达的siRNA(TR-siRNA,TERT-siRNA),通过脂质体法将其转染膀胱移行细胞癌BIU-87细胞株,采用荧光定量PCR分析TR、TERT mRNA表达,TRAP-ELISA检测端粒酶活性,MTT法检测细胞增殖。结果3对TERT-siRNA和3对TR-siRNA中,pRNAT-TERT-Ⅲ和pRNAT-TR-Ⅲ可以特异性抑制BIU-87细胞株中TERT和TR表达(分别减少67%和41%,P<0.05),且BIU-87细胞株的生长和端粒酶活性明显受到抑制。结论TR-siRNA和TERT-siRNA能特异且高效阻断各自基因表达,降低端粒酶活性,进而抑制BIU-87细胞的增殖。Objective Telomerase activation is correlated to genesis, progression,invasion and metastasis of transitional cell carcinoma of bladder. This study was to evaluate the effects of small interfering RNA （ siRNA ） against either the template RNA （human telomerase RNA, hTR ） or its catalytic subunit （human telomerase reverse transcriptase, hTERT ） of telomerase gene on the telomerase activity and the growth inhibition of human transitional cell carcinoma of bladder cell line BIU - 87. Methods Three human TR - specific small interfering RNAs （ TR - siRNA ） and three human TERT - specific small interfering RNAs （ TERT - siRNA ） were designed based on TR and TERT mRNA sequences. TR - siRNA and TERT - siRNA were transfeeted into transitional cell carcinoma of bladder cell line BIU - 87 by using liposome transfection. The influence on the TR mRNA and TERT mRNA were analyzed by fluorescence quantitative PCR, while telomeric repeat amplification protocol（TRAP ） was applied to detect the telomerase activity, and the growth inhibition of BIU -87 ceils was detected by MTT assay. Results Among 3 pairs of TERT - siRNA and 3 pairs of TR - siRNA, pRNAT - TERT - Ⅲ and pRNAT - TR - Ⅲ could significantly reduce the expression of TERT and TR mRNA in BIU - 87 cells by 63% and 41% （ P 〈 0.05 ） , the growth of BIU - 87 cells were inhibited and telomerase activity were considerably decreased. Conclusions TR - siRNA and TERT - siRNA can specifically and effectively reduce both TR and TERT mRNA expression, inhibit the oroliferation by bloeking telomerase activity of BIU - 87 cells.

关 键 词：小干扰RNA 端粒酶RNA 端粒酶逆转录酶 膀胱移行细胞癌 

分 类 号：R737.14[医药卫生—肿瘤]