miR-497-5p对人宫颈癌细胞增殖的抑制作用及机制

发布时间：2018-05-28 18:20

本文选题：子宫颈癌 + 微小RNA--p　；参考：《山东医药》2017年05期

【摘要】：目的探讨miR-497-5p对人宫颈癌细胞(He La)增殖的抑制作用及机制。方法将对数生长期人宫颈癌He La细胞随机分为两部分,第一部分细胞随机分为四组:miR-497-5p mimics NC+IKCa1 3'端非翻译区(3'-URT)野生型双荧光素酶重组质粒载体(IKCa1-wt)转染组、miR-497-5p mimics+IKCa1-wt转染组、miR-497-5p mimics NC+3'-URT突变型双荧光素酶重组质粒载体(IKCa1-mut)转染组、miR-497-5p mimics+IKCa1-mut转染组,分别转染相应质粒48 h,用双荧光素酶报告基因检测系统检测He La细胞荧光素酶活性。第二部分细胞随机分为三组:miR-497-5p mimics转染组、miR-497-5p mimics NC转染组和空白对照组,分别转染相应质粒,用qRT-PCR技术检测细胞IKCa1 mRNA表达,Western boltting法检测细胞IKCa1蛋白表达,CCK-8法检测细胞增殖能力。结果第一部分细胞中的miR-497-5p mimics+IKCa1-wt转染组荧光素酶活性较其他三组降低(P均0.05),其他三组间比较差异无统计学意义(P均0.05),证实miR-497-5p与IKCa1 3'-URT存在特异性结合位点。第二部分细胞中的miR-497-5p mimics转染组IKCa1 mRNA、蛋白的相对表达量较其他两组降低(P均0.05),其他两组比较差异无统计学意义(P均0.05)。第二部分细胞培养24、48、72、96 h,miR-497-5p mimics转染组增殖能力均较其他两组降低(P均0.05),其他两组比较差异无统计学意义(P均0.05)。结论 miR-497-5p可以通过负向调控IKCa1基因表达以抑制宫颈癌细胞增殖。
[Abstract]:Objective to investigate the inhibitory effect and mechanism of miR-497-5p on the proliferation of human cervical cancer cell line he La. Methods Human cervical cancer He-La cells in logarithmic phase were randomly divided into two parts. The first part was randomly divided into four groups: MiR-497-5p mimics NC IKCa1 3'untranslated region 3'-URT) wild-type double-luciferase recombinant plasmid vector IkCa1-wt) transfection group (miR-497-5p mimics IKCa1-wt transfection group) and miR-497-5p mimics NC 3'-URT mutant vector IKCa1-mutated group (MIR-497-5p mimics IKCa1-mut transfection group). The luciferase activity of He-La cells was detected by double luciferase reporter gene detection system at 48 h after transfection of corresponding plasmids. The second part of cells were randomly divided into three groups: 1 miR-497-5p mimics transfection group (n = 10) and control group (n = 10). The corresponding plasmids were transfected respectively. The expression of IKCa1 protein was detected by qRT-PCR technique and the expression of IKCa1 protein was detected by CCK-8 method. Results the luciferase activity in the miR-497-5p mimics IKCa1-wt transfection group was lower than that in the other three groups (P < 0.05), but there was no significant difference among the other three groups (P < 0.05), which confirmed the existence of specific binding sites between miR-497-5p and IKCa1 3'-URT. In the second part, the relative expression of IKCa1 mRNAs in the miR-497-5p mimics transfection group was lower than that in the other two groups (P 0.05), but there was no significant difference between the other two groups (P 0.05). In the second part, the proliferation ability of the mimics transfected group was significantly lower than that of the other two groups, but there was no significant difference between the other two groups. Conclusion miR-497-5p can inhibit the proliferation of cervical cancer cells by negatively regulating the expression of IKCa1 gene.
【作者单位】：西南医科大学附属医院;
【基金】：四川省卫生厅科研基金资助项目(110373) 四川省泸州市科技局科研项目(2013LZLY-J30)
【分类号】：R737.33

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