miR－873靶向生存蛋白基因对人前列腺癌PC3细胞侵袭性的影响

摘要

Objective To identify the role of miR-873, which may regulate the expression of survivin,in human pros-tate cancer PC3 cells invasion.Methods PC3 cells were cultured in vitro, and changes of cellular morphology were ob-served by phase contrast microscope.miR-873,which might regulate the expression of survivin,was predicted by bioinforma-tics and identified using dual luciferase report system.Expressions of miR-873 and survivin were determined using real-time quantitative PCR( qRT-PCR) and Western blotting after transfection of miR-873 mimics.The invasion of PC3 cells was de-tected in vitro by Transwell chamber.Results The expression of survivin was positive by immunofluorescence cytochemis-try.Using dual luciferase reporter system, miR-873 could inhibit the expression of survivin by binding to its mRNA 3′UTR.Results of qRT-PCR and Western blotting showed that overexpression of miR-873 down-regulated the expression of survivin.The invasion of PC3 cells was suppressed by over-expression of miR-873.Conclusion MiR-873 may negatively regulate the expression of survivin in human prostate cancer PC3 cells and inhibit cell invasion.%目的：探讨人前列腺癌PC3细胞中miR－873和生存蛋白（ Survivin）基因的表达，阐明miR－873对生存蛋白基因的靶向调控作用以及对PC3细胞侵袭性的影响。方法培养PC3细胞并应用免疫荧光化学技术检测生存蛋白基因的表达；采用生物信息学方法对miR－873和生存蛋白基因的匹配关系进行预测，并应用双荧光素酶报告基因系统鉴定；转染miR－873模拟物后，行实时定量PCR（ qRT－PCR）检测miR－873和生存蛋白基因的表达，Western印迹检测生存蛋白的表达，Transwell小室检测癌细胞体外的侵袭性。结果倒置相差显微镜下可见细胞均匀分布，形态规则均一；免疫荧光化学显示，胞质内有生存蛋白的表达；靶基因预测软件miRanda和TargetScan显示， miR－873和生存蛋白基因匹配良好，双荧光素酶报告基因系统鉴定发现，miR－873能够结合生存蛋白 mRNA 3′UTR并有效抑制其表达。 qRT－PCR和Western印迹检测结果均表明， miR－873的过表达能够下调生存蛋白基因表达。Transwell实验显示，miR－873的过表达能够抑制PC3细胞的侵袭。结论 miR－873通过靶向作用于生存蛋白mRNA 3′UTR负性调控人前列腺癌PC3细胞中生存蛋白的表达，并能够抑制其侵袭。