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目的:探讨RNA干扰对宫颈癌Hela细胞人端粒酶逆转录酶基因的抑制效应。方法:化学合成靶向于hTERT基因的4个siRNA片段和1个阴性对照小干扰RNA(small interference RNA,siRNA)片段,分siRNA 1-4、control siRNA和空白对照组共6个组,分别用阳离子脂质体转染法将其导入Hela细胞内,通过RT-PCR和WesternBlot方法分别检测宫颈癌细胞转染后细胞hTERT mRNA和蛋白水平,MTS法检测细胞生长增殖,流式细胞仪检测细胞凋亡情况。结果:siRNA-3组端粒酶活性明显下降,hTERT mRNA表达水平明显下降,hTERT蛋白表达明显下降,细胞凋亡率明显升高,与空白对照组比较差异有统计学意义(P<0.01),阴性对照组与空白对照组差异无统计学意义(P>0.05)。结论:靶向于hTERT的siRNA能特异性抑制宫颈癌Hela细胞hTERT基因,下调hTERT mRNA及蛋白表达水平,抑制癌细胞生长增殖,促进宫颈癌细胞凋亡,为宫颈癌的基因治疗研究提供实验依据。
Objective: To investigate the inhibitory effect of RNA interference on human telomerase reverse transcriptase gene in cervical cancer Hela cells. Methods: Four siRNA fragments targeted to hTERT gene and one negative control siRNA (small interfering RNA) fragment were chemically synthesized and divided into 6 groups: siRNA 1-4, control siRNA and blank control group. The expression of hTERT mRNA and protein in cervical cancer cells was detected by RT-PCR and Western Blot. The proliferation and proliferation of cervical cancer cells were detected by MTS and flow cytometry Death situation. Results: The telomerase activity of siRNA-3 group was significantly decreased, the expression of hTERT mRNA was significantly decreased, the expression of hTERT protein was significantly decreased, and the apoptosis rate was significantly increased compared with the control group (P <0.01) There was no significant difference between negative control group and blank control group (P> 0.05). CONCLUSIONS: siRNA targeting hTERT can specifically inhibit the hTERT gene in cervical cancer Hela cells, down-regulate the expression of hTERT mRNA and protein, inhibit the growth and proliferation of cervical cancer cells and provide the experimental evidence for the gene therapy of cervical cancer .