siRNA靶向沉默hTERT基因对宫颈癌Caski细胞特性的影响

发布时间：2018-07-10 00:06

本文选题：宫颈癌 + hTERT基因　；参考：《中国老年学杂志》2017年17期

【摘要】：目的研究siRNA靶向沉默h TERT基因后对宫颈癌Caski细胞特性的影响。方法设计、合成特异性针对h TERT mRNA的siRNA,将其克隆入p Genesil-1.1质粒中,重组成h TERT mRNA-siRNA的表达载体,导致目的基因沉默,是由电转染入宫颈癌Caski细胞,从而产生RNA干扰作用。应用Western印迹法及RT-PCR法检测沉默h TERT基因后对于宫颈癌Caski细胞的蛋白及mRNA的表达情况;检测细胞周期运用流式细胞仪;沉默h TERT基因后宫颈癌Caski细胞的增殖能力可通过CCK-8增殖实验检测。沉默h TERT基因后宫颈癌Caski细胞的侵袭能力可通过小室侵袭实验检测。结果 h TERT基因沉默48 h以后,与空白组及未转染组相比,转染组宫颈癌Caski细胞的蛋白及mRNA表达出现明显降低(P0.05);细胞周期的检测结果显示S期的细胞数目明显减低,转染组的宫颈癌Caski细胞停留于G0/G1周期,空白组、转染组及未转染组相比差异显著(P0.05);宫颈癌Caski细胞在转染组的增殖过程中被明显抑制(P0.05);转染组穿过滤膜的细胞数量明显减少(P0.05)。结论 Si RNA靶向沉默h TERT基因电转染后,使宫颈癌Caski细胞的增殖和迁移能力得到抑制,一定程度上改善了宫颈癌患者的预后。
[Abstract]:Objective to study the effect of hTERT gene silencing by siRNA on the characteristics of cervical cancer Caski cells. Methods siRNAs specifically targeting hTERT mRNA were synthesized and cloned into pGenesil-1.1 plasmid, which was recombined into the expression vector of hTERT mRNA-siRNA, resulting in the silencing of the target gene, which was induced by electrotransfection into cervical cancer Caski cells, resulting in RNA interference. Western blot and RT-PCR were used to detect the expression of protein and mRNA in human cervical cancer Caski cells after hTERT gene silencing, and flow cytometry was used to detect the cell cycle. The proliferative ability of cervical cancer Caski cells after hTERT gene silencing can be detected by CCK-8 proliferative assay. The invasive ability of cervical cancer Caski cells after hTERT gene silencing can be detected by chamber invasion assay. Results after 48 h of hTERT gene silencing, the expression of protein and mRNA of cervical cancer Caski cells in transfected group was significantly lower than that in blank group and untransfected group (P0.05), and the cell cycle detection showed that the number of S phase cells was significantly decreased. In the transfected group, the number of cervical cancer Caski cells was significantly inhibited in the transfected group (P0.05), while in the blank group, the transfected group and the untransfected group were significantly inhibited (P0.05); the number of cells passing through the filter membrane was significantly decreased in the transfected group (P0.05). Conclusion Si RNA targeted silencing hTERT gene can inhibit the proliferation and migration of cervical cancer Caski cells, and improve the prognosis of cervical cancer patients to some extent.
【作者单位】：天津市静海县医院妇产科;天津医科大学总医院;
【分类号】：R737.33

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