山东大学学报(医学版) ›› 2015, Vol. 53 ›› Issue (6): 23-27.doi: 10.6040/j.issn.1671-7554.0.2015.083
曾仁仁, 张俊华, 张银旭
ZENG Renren, ZHANG Junhua, ZHANG Yinxu
摘要: 目的 探讨miR-485-5p对人结直肠癌HCT116细胞中Survivin基因的靶向调控作用以及对细胞侵袭性的影响。方法 培养HCT116细胞,应用免疫荧光检测Survivin基因在细胞中的表达。采用生物信息学方法对miR-485-5p和Survivin基因的靶向匹配关系进行预测,并应用双荧光素酶报告基因系统鉴定。脂质体2000转染miR-485-5p模拟物后,Real-time PCR检测miR-485-5p和Survivin基因的表达,Western blotting检测Survivin蛋白的表达,Transwell小室检测HCT116细胞体外的侵袭性。结果 倒置相差显微镜下可见细胞均匀分布,形态规则均一;细胞免疫荧光显示胞质内有Survivin蛋白的表达。靶基因预测软件miRanda和TargetScan显示miR-485-5p和Survivin基因匹配良好,双荧光素酶报告基因系统鉴定发现miR-485-5p能够结合Survivin mRNA 3'UTR,并有效抑制其表达。Real-time PCR和Western blotting检测结果显示,过表达miR-485-5p能够导致Survivin基因表达的下降。Transwell实验结果显示,miR-485-5p的过表达能够抑制HCT116细胞的侵袭。结论 miR-485-5p通过靶向作用于Survivin mRNA 3'UTR能够负性调控Survivin基因的表达,并能够抑制人结直肠癌HCT116细胞的侵袭。
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