【摘 要】
：
目的:研究miR-185-5p抑制支气管上皮细胞16HBE增殖的调控作用及其机制. 方法:用实时荧光定量PCR(qRT-PCR)检测了A549和16HBE细胞内miR-185-5p和PLAC8 mRNA的表达量,用Lipofectamine 3000 转染miR-185-5p mimics到16HBE细胞中,用双荧光素酶活性实验检测了 miR-185-5p 对 PLAC8 的靶向性.用 MTT检测了miR-185-5p对16HBE细胞增殖的影响,用流式细胞术检测了细胞周期分布,用 Western blot
【机 构】
：
中南民族大学 生命科学学院 & 医学生物研究所 &武陵山区特色资源植物种质保护与利用湖北省重点实验室,武汉430074
论文部分内容阅读
目的:研究miR-185-5p抑制支气管上皮细胞16HBE增殖的调控作用及其机制. 方法:用实时荧光定量PCR(qRT-PCR)检测了A549和16HBE细胞内miR-185-5p和PLAC8 mRNA的表达量,用Lipofectamine 3000 转染miR-185-5p mimics到16HBE细胞中,用双荧光素酶活性实验检测了 miR-185-5p 对 PLAC8 的靶向性.用 MTT检测了miR-185-5p对16HBE细胞增殖的影响,用流式细胞术检测了细胞周期分布,用 Western blotting检测了细胞内蛋白表达水平. 结果:miR-185-5p在正常人肺支气管上皮细胞16HBE中的表达水平低于肺癌细胞A549(P<0. 05);将16HBE细胞转染不同浓度的miR-185-5p mimics,16HBE细胞的增殖活性随着miR-185-5p mimics浓度升高而下降(P<0. 05). 通过生物信息学预测PLAC8可能是miR-185-5p的一个靶基因. 16HBE细胞中PLAC8的表达量高于A549(P<0. 05).与转染NC对照组比较,转染不同浓度梯度的miR-185-5p mimics到16HBE细胞后48 h,细胞内PLAC8 mRNA含量和蛋白表达量明显降低(P<0. 05).双荧光素酶活性结果表明miR-185-5p 与 PLAC8 3′UTR结合后抑制其表达. miR-185-5p使细胞周期阻滞在S期,抑制了细胞内c-Myc、CDK2、CyclinA2、P21、p-Cdc2蛋白的表达.结论:miR-185-5p可能通过抑制 PLAC8基因表达,抑制16HBE细胞的增殖活性.
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