沉默INK4基因座中反义非编码RNA对人宫颈癌细胞株HeLa增殖的影响及其机制

发布时间：2018-05-12 05:08

本文选题：宫颈肿瘤 + 宫颈癌　；参考：《山东医药》2017年35期

【摘要】：目的观察沉默INK4基因座中反义非编码RNA(ANRILA)对人宫颈癌细胞株He La增殖的影响,并探讨其机制。方法取对数生长期He La细胞分为观察组和对照组,分别ANRIL-siRNA、对照siRNA乱序。转染24、48 h时采用CCK-8法观察两组细胞增殖情况(结果以OD450表示)。转染48 h时采用荧光定量PCR法检测两组细胞ANRIL、p21、周期蛋白依赖性激酶6(CDK6)、周期蛋白依赖性激酶2(CDK2)mRNA相对表达量,采用CHIP实验观察两组细胞p21基因H3K27位点三甲基化状态。结果转染24 h观察组、对照组OD450分别为0.55±0.08、0.84±0.11,转染72 h观察组、对照组OD450分别为1.21±0.12、1.82±0.14。转染24、72 h观察组OD450均低于对照组(P0.05)。转染48 h观察组ANRIL及CDK6、CDK2 mRNA相对表达量低于对照组(P均0.05),p21 mRNA相对表达量高于对照组(P均0.05)。转染48 h两组细胞p21基因H3K27位点三甲基化水平观察组、对照组分别为1.19±0.08、5.62±0.21,观察组低于对照组(P0.05)。结论沉默ANRIL可抑制宫颈癌He La细胞的增殖。其机制是沉默ANRIL可以降低宫颈癌He La细胞p21基因H3K27位点三甲基化水平,促进p21mRNA表达,抑制CDK6、CDK2 mRNA表达,从而抑制宫颈癌He La细胞增殖。
[Abstract]:Objective to investigate the effect of antisense noncoding RNAs ANRILA in silencing INK4 locus on the proliferation of human cervical cancer cell line he La and its mechanism. Methods he La cells in logarithmic growth phase were divided into two groups: observation group and control group. ANRIL-siRNAs and control siRNA were randomly divided into two groups. The proliferation of two groups of cells was observed by CCK-8 method at 48 h after transfection. The results were expressed as OD450. At 48 h after transfection, the relative expression of ANRIL p21, cyclin dependent kinase 6 CDK6 and cyclin dependent kinase 2(CDK2)mRNA was detected by fluorescence quantitative PCR assay. The trimethylation of H3K27 site of p21 gene in two groups was observed by CHIP assay. Results after 24 h transfection, the OD450 of the control group was 0.55 卤0.08 卤0.84 卤0.11, and the OD450 of the control group was 1.21 卤0.121.82 卤0.14 after 72 h transfection. The OD450 of the observation group was lower than that of the control group for 72 h. The relative expression of ANRIL and CDK6 + CDK2 mRNA in the observation group was lower than that in the control group at 48 h after transfection. The relative expression of p21 mRNA in the control group was higher than that in the control group (P < 0.05). The level of H3K27 trimethylation of p21 gene was 1.19 卤0.08c5.62 卤0.21 in the control group, which was lower than that in the control group (P 0.05) at 48 h after transfection. Conclusion silencing ANRIL can inhibit the proliferation of helium La cells. The mechanism is that silencing ANRIL can reduce the level of H3K27 trimethylation of p21 gene, promote the expression of p21mRNA and inhibit the expression of CDK6 + CDK2 mRNA in cervical cancer cells, thus inhibit the proliferation of He-La cells.
【作者单位】：黔西南州人民医院;
【分类号】：R737.33

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