RNA干扰沉默人端粒酶逆转录酶基因对胶质瘤U87细胞增殖及凋亡的影响  

The effect of silencing human telomerase reverse transcriptase by RNA interference on U87 glioma cell proliferation and apoptosis

作　　者：陈健[1] 陈谦学[1] 

机构地区：[1]武汉大学人民医院神经外科 430060

出　　处：《中华实验外科杂志》2014年第9期1868-1870,共3页Chinese Journal of Experimental Surgery

摘　　要：目的 观察人端粒酶逆转录酶（hTERT）对胶质瘤U87细胞增殖及凋亡的影响.方法 构建靶向hTERT的小干扰RNA（siRNA）表达质粒转染U87细胞,通过Western blot检测hTERT蛋白的表达水平,通过逆转录-聚合酶链反应（RT-PCR）检测hTERT mRNA表达水平,采用噻唑蓝（MTT）法及流式细胞仪分别检测转染后U87细胞的增殖活性及凋亡率.结果 与空白对照组及阴性对照组比较,各siRNA-hTERT干扰组hTERT蛋白表达水平明显降低;各siRNA-hTERT干扰组hTERT mRNA相对表达量分别为0.79 ±0.11、0.58±0.15、0.39±0.16,与阴性对照组比较明显降低,其差异有统计学意义（P＜0.05）;与空白对照组及阴性对照组比较,转染siRNA-hTERT后U87细胞的增殖受到明显抑制且在第3天时最明显;转染48 h后各siRNA-hTERT干扰组U87细胞凋亡率分别为（14.31±0.93）％、（17.57±0.61）％、（22.68±1.04）％,与空白对照组及阴性对照组比较明显增加,其差异有统计学意义（P＜0.05）.结论 靶向hTERT基因的siRNA转染U87细胞后可以显著抑制细胞增殖并诱导细胞凋亡的发生.Objective To observe the effect of human telomerase reverse transcriptase （hTERT） on U87 glioma cell proliferation and apoptosis.Methods The small interfering RNA （siRNA） expression plasmid targeting hTERT was constructed and transfected into U87 glioma cells.The expression level of hTERT protein and mRNA was detected by Western blotting and reverse transcription-polymerase chain reaction （RT-PCR） respectively.The U87 glioma cell proliferation and apoptosis were analyzed by methyl thiazol tetrazolium （MTT） assay and flow cytometry,respectively.Results Compared with the blank control group and the negative control group,the expression level of hTERT protein in each siRNA group was significantly reduced （P 〈 0.05）.The relative expression level of hTERT mRNA in each siRNA group was 0.79 ±0.11,0.58 ±0.15 and 0.39 ±0.16 respectively,which was significantly lower than in the negative control group （P 〈 0.05）.As compared with the blank control group and the negative control group,the proliferation of U87 glioma cells after transfection was significantly inhibited and the strongest inhibitory effect was observed on the third day.The apoptosis rate of U87 glioma cells of in each siRNA group which was transfected after 48 h was （14.31 ± 0.93） %,（17.57 ± 0.61） % and （22.68 ± 1.04） % respectively,which was significantly higher than in the blank control group and the negative control group （P 〈0.05）.Conclusion The silence of hTERT gene after siRNA targeting hTERT being transfected into U87 glioma cells can significantly inhibit cell proliferation and induce cell apoptosis.

关 键 词：RNA干扰 人端粒酶逆转录酶 胶质瘤 细胞增殖 

分 类 号：R739.41[医药卫生—肿瘤]