长链非编码RNA 565686通过靶向调控微小RNA-195对前列腺癌细胞增殖、迁移和侵袭的影响

Long non-coding RNA 565686 regulates the proliferation, migration and invasion of prostate cancer cells by targeting microRNA-195

目的:探讨长链非编码RNA 565686(lncRNA-565686)对前列腺癌细胞增殖、迁移和侵袭的影响及其分子机制。方法:收集武汉大学人民医院2017年1月至2020年1月行前列腺癌根治术的33例前列腺癌组织和同期行经尿道前列腺电切术的45例良性前列腺增生组织标本,采用实时定量反转录聚合酶链反应(RT-qPCR)技术检测lncRNA-565686在前列腺组织和前列腺癌细胞的相对表达水平。在前列腺癌细胞(PC-3)中转染lncRNA-565686短发卡RNA,分别将未转染短发卡RNA、转染空载体和转染短发卡RNA细胞分为未转染组、空载体转染组和转染组。分别采用细胞克隆增殖实验检测细胞增殖能力;采用细胞迁移和侵袭实验(Transwell实验)检测细胞迁移和侵袭能力;采用双荧光素酶报告基因实验验证lncRNA-565686与微小RNA-195(miR-195)的靶向调控关系。组间比较采用t检验。结果:前列腺癌组织中lncRNA-565686的相对表达水平高于良性前列腺增生组织,差异有统计学意义(2.51±0.19比0.99±0.13,t=39.826,P<0.05);前列腺癌淋巴结细胞(LNCaP)、PC-3细胞中lncRNA-565686表达水平高于人正常前列腺上皮细胞(RWPE-1)细胞,差异有统计学意义(2.36±0.04和3.58±0.06比0.99±0.06,t=40.257、68.757,P<0.05);干扰lncRNA-565686表达后,转染组细胞增殖、迁移和侵袭能力均低于未转染组和空载体转染组。双荧光素酶报告基因实验证实lncRNA-565686与miR-195之间的直接调控关系。结论:lncRNA-565686在前列腺癌中高表达,下调lncRNA-565686的表达可明显抑制前列腺癌细胞的增殖、迁移和侵袭能力,其机制可能与直接负向调控miR-195的表达相关。

Objective To investigate the effect of long non-coding RNA 565686(lncRNA-565686)on the proliferation,migration and invasion of prostate cancer cells and the potential molecular mechanism.Methods Totally,33 cases of prostate cancer and 45 cases of benign prostate tissues from January 2015 to January 2020 were collected,and the relative expression level of lncRNA-565686 in prostate cancer tissues and cell lines was detected by real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR).The lncRNA-565686 short hairpin RNA(shRNA)was transfected into prostate cancer cell line(PC-3)cells as untransfected group,and empty vector transfected group,and transfected group were set up.The cell proliferation ability was detected by cell cloning and proliferation assay,and cell migration and invasion ability was detected by Transwell assay.The dual luciferase reporter gene experiment confirmed the targeted regulation between lncRNA-565686 and microRNA-195(miR-195).The t test was used for comparison between groups.Results The relative expression level of lncRNA-565686 in prostate cancer tissues was significantly higher than in adjacent tissues(2.51±0.19 vs.0.99±0.13,t=39.826,P<0.05).The expression level of lncRNA-565686 in lymph node carcinoma of the prostate(LNCaP)and PC-3 cells was significantly higher than in human prostate epithelial cells(RWPE-1)cells,and the difference was statistically significant(2.36±0.04 and 3.58±0.06 vs.0.99±0.06,t=40.257,68.757,P<0.05).After transfection with shRNA,the proliferation,migration and invasion ability of cells in the transfected group was significantly decreased as compared with that in the untransfected group and the empty vector transfected group.Dual luciferase reporter gene assay confirmed the direct regulatory relationship between lncRNA-565686 and miR-195.Conclusion LncRNA-565686 is highly expressed in prostate cancer,and negatively regulates miR-195 to affect the prostate cancer cells.