RNA干扰人端粒酶hTR基因对HeLa细胞增殖的影响

Effects of inhibition of human telomerase RNA by RNA interference on proliferation of HeLa cells

目的:探讨RNA干扰抑制人端粒酶RNA组分(human telomerase RNA component,hTR)的表达对宫颈癌细胞(HeLa细胞)增殖的影响。方法:hTR基因的RNA干扰表达重组体用脂质体介导方法稳定转染HeLa细胞。RT-PCR法检测hTR的mRNA表达水平,TRAP-PCR法检测端粒酶活性,MTT法检测细胞增殖。结果:RT-PCR结果表明,稳定转染RNA干扰表达重组体的细胞株,其hTR基因的mRNA抑制率较无转染对照组下降了(59.7±3.3)%,较转染空载体组下降了(56.3±4.4)%;实验组细胞端粒酶活性下降,生长速度明显减慢。结论:利用RNA干扰技术抑制hTR基因在HeLa细胞的表达能抑制该细胞端粒酶活性和增殖活性。

Objective To investigate the effect of inhibiting hTR expression by RNA interference on the proliferation of HeLa cell. Methods A combination plasmids expressing shRNA specific for human telomerase RNA were transfected by Lipofectamine into HeLa cells. The inhibitory of hTR mRNA level was assayed by RT-PCR. The activity of telomerase was tested by TRAP-EB staining. The proliferation activity of HeLa cells was determined using methyl thiazolyl tetrazolium （MTT）. Results As detected by RT-PCR,mRNA level of hTR in HeLa cells transfected by shRNA was reduced （59.7 ± 3.3）% and （56.3 ± 4.4）% than that of untransfected control and vector control,respectively. Telomerase activity of HeLa cells transfected by shRNA was decreased.The proliferation of HeLa cells transfected by shRNA was significantly reduced. Conclusion shRNA of human telomerase RNA can inhibit the expression of hTR and subsequently inhibit telomerase activity and the proliferation of HeLa cells.