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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
  • ISSN号:1000-3282
  • 期刊名称:《生物化学与生物物理进展》
  • 时间:0
  • 分类:R737.14[医药卫生—肿瘤;医药卫生—临床医学]
  • 作者机构:[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
  • 相关基金:基金项目:国家自然科学基金(30572030).
中文摘要:

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。

英文摘要:

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.

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期刊信息
  • 《生物化学与生物物理进展》
  • 中国科技核心期刊
  • 主管单位:中国科学院
  • 主办单位:中国科学院生物物理研究所 中国生物物理学会
  • 主编:王大成
  • 地址:北京市朝阳区大屯路15号
  • 邮编:100101
  • 邮箱:prog@sun5.ibp.ac.cn
  • 电话:010-64888459
  • 国际标准刊号:ISSN:1000-3282
  • 国内统一刊号:ISSN:11-2161/Q
  • 邮发代号:2-816
  • 获奖情况:
  • 1999年中国期刊奖提名奖,2000年中国科学院优秀期刊特别奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),荷兰文摘与引文数据库,美国剑桥科学文摘,美国科学引文索引(扩展库),美国生物科学数据库,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:18821