miR-142-5p靶向调控TOP2A基因对前列腺癌细胞增殖和侵袭的影响  

Effects of miR-142-5p targeting TOP2A gene on proliferation and invasion of prostate cancer cells

作　　者：朱延杰[1] 尤平洪[1] 唐良友 段晓波[1] 张姚[1] 代庆德[1] 吕东[2] ZHU Yanjie;YOU Pinghong;TANG Liangyou;DUAN Xiaobo;ZHANG Yao;DAI Qingde;LYU Dong(People's Hospital of Deyang City,Deyang 618000,China)

机构地区：[1]德阳市人民医院,四川德阳618000 [2]四川省人民医院(东院)

出　　处：《山东医药》2020年第14期13-16,共4页Shandong Medical Journal

基　　金：四川省卫生计生委科研课题资助项目(16PJ437)。

摘　　要：目的观察miR-142-5p靶向调控拓扑异构酶Ⅱα(TOP2A)基因对前列腺癌细胞增殖和侵袭的影响。方法收集前列腺癌组织和相应的癌旁组织40例,采用qPCR方法检测miR-142-5p、TOP2A mRNA表达,Pearson等级相关分析前列腺癌组织中miR-142-5p与TOP2A表达的相关性。常规培养前列腺癌LNCaP细胞,应用TargetScan7.1预测软件及双荧光素酶报道基因试验判断miR-142-5p是否对TOP2A基因具有靶向调控作用。将LNCaP细胞分为正常对照组、转染对照组和TOP2A过表达组,正常对照组转染NC质粒,转染对照组转染miR-142-5p mimic,TOP2A过表达组同时转染miR-142-5p mimic和TOP2A过表达质粒。采用MTS实验检测细胞增殖能力,Boyden实验检测细胞侵袭能力,Western blotting法检测细胞中Wnt/β-catenin信号通路的活性。结果与癌旁组织比较,前列腺癌组织中miR-142-5p表达降低、TOP2A基因表达升高(P均<0.05),二者表达呈负相关(r=-0.601,P<0.05)。TargetScan7.1软件在线预测显示,TOP2A启动子区与miR-142-5p具有结合位点;双荧光素酶报告基因试验结果显示,与NC wt组相比,miR-142-5p wt组荧光素酶活性降低(P<0.05);而NC mut组与miR-142-5p mut组间荧光素酶活性无统计学差异(P>0.05)。与正常对照组相比,转染对照组和TOP2A过表达组细胞增殖率降低,穿膜细胞数减少(P均<0.05);与转染对照组相比,TOP2A过表达组细胞增殖率升高,穿膜细胞数增加(P均<0.05)。与正常对照组相比,转染对照组和TOP2A过表达组细胞中Wnt3a、β-catenin蛋白表达降低;与转染对照组相比,TOP2A过表达组细胞中Wnt3a、β-catenin蛋白表达增加(P均<0.05)。结论前列腺癌组织中miR-142-5p表达降低,TOP2A表达增加,二者呈负相关;miR-142-5p能够靶向调控TOP2A表达,从而抑制前列腺癌细胞的增殖与侵袭能力。Objective To observe the effects of miR-142-5p targeting topoisomeraseⅡα(TOP2A)gene on the proliferation and invasion of prostate cancer cells.Methods Forty cases of prostate cancer tissues and corresponding adjacent tissues were collected,miR-142-5p and TOP2A mRNA expression was detected by qPCR.LNCaP cells of prostate cancer were routinely cultured.TargetScan7.1 prediction software and dual luciferase reporter gene test were used to detect miR-142-5p targeted regulation of TOP2A gene.LNCaP cells were divided into the normal control group,the transfection control group and the TOP2A overexpression group.The LNCaP cells in the normal control group were transfected with the NC plasmid,the transfection control group with the miR-142-5p mimic,and the TOP2A overexpression group with the miR-142-5p mimic and the TOP2A overexpression plasmid.MTS test was used to detect cell proliferation ability,Boyden test was used to detect cell invasion ability,and Western blotting was used to detect the activity of Wnt/β-catenin signaling pathway in cells.Results Compared with the adjacent tissues,the expression of miR-142-5p in the prostate cancer tissues decreased,the expression of TOP2A increased,and the two expression levels were negatively correlated(r=-0.601,both P<0.05).Online prediction by TargetScan7.1 showed that the TOP2A promoter region had binding sites with miR-142-5p.The results of double luciferase reporter gene test showed that compared with the NC wt group,the luciferase activity of miR-142-5p wt group decreased(P<0.05).There was no significant difference in luciferase activity between the NC mut group and the miR-142-5p mut group(P>0.05).Compared with the normal control group,the cell proliferation rate of cells in the transfection control group and the TOP2A overexpression group decreased,and the number of transmembrane cells decreased(all P<0.05).Compared with the transfection control group,the cell proliferation rate increased,and the number of transmembrane cells increased in the TOP2A overexpression grou

关 键 词：前列腺癌 miR-142-5p 拓扑异构酶Ⅱα 细胞增殖 细胞侵袭 WNT/Β-CATENIN通路 

分 类 号：R473.73[医药卫生—护理学]