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  

Impact of miR-873 on human prostate cancer PC3 cells invasion

作　　者：李锬[1] 李晨光[1] 刘岩[1] 李炳勋[1] 杨先旭[1] 

机构地区：[1]辽宁医学院附属第一医院泌尿外科,辽宁锦州121001

出　　处：《军事医学》2015年第8期610-613,共4页Military Medical Sciences

摘　　要：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Target Scan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。Objective To identify the role of miR-873, which may regulate the expression of survivin,in human prosrate cancer PC3 cells invasion. Methods PC3 cells were cultured in vitro, and changes of cellular morphology were observed by phase contrast microscope, miR-873, which might regulate the expression of survivin, was predicted by bioinforrnatics 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 transfeetion of miR-873 mimics. The invasion of PC3 cells was detected in vitro by Transwell chamber. Results The expression of survivin was positive by immunofluoreseence cytochemistry. 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.

关 键 词：微小RNA 生存蛋白 前列腺癌PC3细胞 

分 类 号：R737.25[医药卫生—肿瘤]