RNA干扰下调hTERT表达对宫颈癌细胞增殖及侵袭能力的影响  

Down-regulation of h TERT expression by RNA interference inhibits the proliferation and invasion ability of cervical cancer He La cells

作　　者：陆杲川[1] 林晓[1] 施慧静 包姿萍 

机构地区：[1]上海市第八人民医院妇产科,上海200235

出　　处：《同济大学学报（医学版）》2014年第5期33-37,共5页Journal of Tongji University(Medical Science)

基　　金：上海市卫生局科研基金(2006122)

摘　　要：目的观察RNA干扰下调人类端粒酶反转录酶(h TERT)表达对宫颈癌He La细胞增殖及侵袭能力的影响。方法针对h TERT基因设计两对siRNA片段,瞬时转染人宫颈癌He La细胞;采用RT-PCR、Western印迹法检测转染后He La细胞h TERT在mRNA水平和蛋白水平的表达变化;PCR-TRAP法检测细胞端粒酶活性;四甲基偶氮唑蓝(MTT)法检测细胞增殖能力变化;流式细胞仪检测细胞周期和凋亡;transwell实验检测细胞运动侵袭能力。结果 RT-PCR及Western印迹法结果显示,h TERT-siRNA能有效下调h TERT在mRNA和蛋白水平的表达,抑制效率分别为(85.8±3.26)%、(97.5±3.72)%;RNA干扰下调h TERT基因表达后,He La细胞端粒酶活性抑制效率为75.5%,增殖受到明显抑制(P<0.05);流式细胞仪检测结果显示,处于G0/G1期细胞增多,由(58.18±3.76)%上升到(68.97±4.01)%(P<0.05);未处理组细胞凋亡(4.14±0.79)%,h TERT-si-RNA组细胞凋亡(8.1±1.22)%,细胞凋亡率增加(P<0.05)。transwell实验检测结果显示,细胞运动侵袭能力下降约30%(P<0.05)。结论 h TERT-siRNA可有效下调h TERT在人宫颈癌He La细胞中的表达,抑制He La细胞生长并降低其侵袭能力。Objective To investigate the effect of down-regulation of h TERT（ human telomerase reverse transcriptase） by RNA interference on the proliferation and invasion ability of human cervical cancer He La cells. Methods Twopair of siRNA targetingh TERTwere designed and constructed,then transfected intoHe La cells. Real time PCR and Western Blot were used todetect the expression of h TERTmRNA and protein levels, respectively. PCR-TRAP method was employed totest the telomerase activity. M TTmethod was performed toanalyze the proliferation inhibition of cells. Flowcytometry was used todetect the cell cycle and apoptosis. Transwell test was used toevaluate the invasion ability of He La cells. Results Real-Time PCR and Western blottingrevealed that h TERTsiRNA significantly down-regulated h TERTexpression at mRNA and protein levels by（ 85. 8 ±3. 26） % and（97. 5 ± 3. 72） %,respectively; the telomerase activity decreased by 75. 5%,and the proliferation of He La cells was significantly inhibited. The proportion of cells in G0/ G1 phase alsodecreased from（ 58. 18 ± 3. 76） % to（ 68. 97 ± 4. 01） % after RNA interference（ P〈0. 05）.Apoptosis rate was significantly enhanced from（ 4. 14 ± 0. 79） % to（ 8. 1 ± 1. 22） %（ P〈0. 05）.Transwell test showed that the invasion ability of He La cells decreased by 30%（ P〈0. 05）.Conclusion RNA interference targetingh TERTcan down-regulate the expression of h TERT,resulting the inhibition of proliferation and invasion ability of He La cells.

关 键 词：RNA干扰 人类端粒酶反转录酶 宫颈肿瘤 

分 类 号：R373.33[医药卫生—病原生物学]