miRNA-191对前列腺癌的增殖、迁移侵袭能力的影响及其机制

miRNA-191 promotes proliferation,migration and invasion of prostate cancer by targeting PLCD1

目的:观察miRNA-191对前列腺癌的增殖、迁移和侵袭能力的影响,并探讨其机制。方法:分别检测4种人前列癌细胞系(PC-3、DU-145、LNCa P、22RU1)及人正常前列腺细胞RWPE-2中miRNA-191的表达水平,并选择前列腺癌细胞系PC-3作为实验对象。将PC-3细胞分为3组:空白对照组(不转染)、miRNA-191 NC组(Inhibitor NC转染PC-3细胞)、miRNA-191 Inhibitor组(miRNA-191 Inhibitor转染PC-3细胞),每组设置3个复孔。采用RTq PCR法检测PC-3细胞miRNA-191和PLCD1的mRNA表达水平;采用CCK8法检测PC-3细胞增殖水平;采用划痕实验和侵袭实验分别检测PC-3细胞迁移能力和侵袭能力;通过Targetscan靶基因预测网站,筛选PLCD1作为miRNA-191的靶向蛋白,并用双荧光素酶靶标实验验证;采用Western blot法检测PC-3细胞PLCD1的蛋白表达。结果:与RWPE-2细胞相比,人前列癌细胞中mi NRA-191的表达水平显著升高(P<0.05),且miRNA-191的表达水平在PC-3中较其他3种细胞系显著上调(P<0.05)。抑制miRNA-191的表达水平后,PLCD1表达水平显著升高,PC-3细胞增殖能力受到抑制,迁移和侵袭能力较空白对照组和miRNA-191 NC组显著降低(P<0.05)。双荧光素酶报告基因实验结果显示,PLCD1基因是miRNA-191的靶基因。结论:miRNA-191通过靶向PLCD1促进前列腺癌PC-3细胞的增殖、迁移和侵袭能力。

Objective:To investigate the effects of miRNA-191 on the proliferation,migration and invasion of prostate cancer,and to explore its mechanism.Methods:The expression levels of miRNA-191 in four human prostate cancer cell lines(PC-3,DU-145,LNCa P,22RU1)and human normal prostate cell line RWPE-2 were detected,and prostate cancer cell line PC-3 was selected as the experimental object.PC-3 cells were divided into three groups:blank control group(no transfection),miRNA-191 NC group(PC-3 cells transfected with Inhibitor NC)and miRNA-191 Inhibitor group(PC-3 cells transfected with miRNA-191 Inhibitor),and each group was provided with three multiple pores.The expression levels of miRNA-191 and PLCD1 were detected by RT-PCR.The cell proliferation was detected by CCK8 assay.Scratch test and invasive test were used to detect cell migration and invasive ability.Through Targetscan target gene prediction website,PLCD1 was screened as the target protein of miRNA-191,and verified by double luciferase target experiment.Western blot assay was used to detect the expression of PLCD1 in cells of each group.Results:Compared with RWPE-2 cells,the expression level of miRNA-191 in human prostate cancer cells was significantly higher(P<0.05),and the expression level of miRNA191 in PC-3 was significantly higher than that in other three cell lines(P<0.05).After inhibiting the expression of miRNA-191,the expression levels of PLCD1 was significantly higher while PC-3 cells'proliferation ability was inhibited,and their migration and invasion ability were significantly lower than those of blank control group and miRNA-191 NC group(P<0.05).The results of double luciferase reporter gene assay showed that PLCD1 gene was a target gene of miRNA-191.Conclusion:miRNA-191 promote the proliferation,migration and invasion of prostate cancer PC-3 cells by targeting PLCD1.