CAJ | 학술논문

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。
목적：탐토인전렬선암PC3세포중miR－873화생존단백（ Survivin）기인적표체，천명miR－873대생존단백기인적파향조공작용이급대PC3세포침습성적영향。방법배양PC3세포병응용면역형광화학기술검측생존단백기인적표체；채용생물신식학방법대miR－873화생존단백기인적필배관계진행예측，병응용쌍형광소매보고기인계통감정；전염miR－873모의물후，행실시정량PCR（ qRT－PCR）검측miR－873화생존단백기인적표체，Western인적검측생존단백적표체，Transwell소실검측암세포체외적침습성。결과도치상차현미경하가견세포균균분포，형태규칙균일；면역형광화학현시，포질내유생존단백적표체；파기인예측연건miRanda화TargetScan현시， miR－873화생존단백기인필배량호，쌍형광소매보고기인계통감정발현，miR－873능구결합생존단백 mRNA 3′UTR병유효억제기표체。 qRT－PCR화Western인적검측결과균표명， miR－873적과표체능구하조생존단백기인표체。Transwell실험현시，miR－873적과표체능구억제PC3세포적침습。결론 miR－873통과파향작용우생존단백mRNA 3′UTR부성조공인전렬선암PC3세포중생존단백적표체，병능구억제기침습。
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.