miR-3064-5p对前列腺癌PC3细胞的作用及相关机制

The Effect and Mechanism of miRNA-3064-5p on Prostate Cancer PC3 Cell Line

目的探讨miR-3064-5p在前列腺癌(PCa)细胞增殖和转移中的作用和潜在机制。方法利用qRT-PCR检测前列腺正常上皮细胞及PCa细胞中miR-3064-5p的表达。脂质体法将阴性对照(mimic nc)及miR-3064-5p模拟物(miR-3064-5p mimic)分别转染入PCa细胞PC3中,细胞克隆形成、MTT实验、划痕愈合、Transwell实验分别检测PC3细胞增殖和迁移侵袭能力,双荧光素酶基因报告法检测miR-3064-5p与真核翻译起始因子5(EIF5)的靶向关系,Western blot检测各组细胞EIF5的表达。结果与正常前列腺上皮细胞相比,PCa细胞中miR-3064-5p的表达降低(P<0.01);过表达miR-3064-5p可抑制PC3细胞的增殖和转移(P均<0.05);生物信息学分析显示,EIF5是miR-3064-5p的一个潜在作用靶点,双荧光素酶报告法证实miR-3064-5p与EIF5基因间存在结合位点,且过表达miR-3064-5p可降低PC3细胞中EIF5的蛋白表达。结论miR-3064-5p在PCa细胞中低表达,其可能通过靶向调控EIF5基因的表达抑制PCa细胞的增殖和转移。

Objective To explore the role and potential mechanism of miRNA-3064-5p in the proliferation,migration and invasion of prostate cancer(PCa)cells.Methods qRT-PCR was used to detect the expression of miR-3064-5p in normal prostate epithelial cells and prostate cancer cells.The liposome method was used to transfect miRNA-3064-5p mimic(miR-3064-5p mimic)and its non-target control(mimic nc)into prostate cancer cells PC3,respectively,and then cell colony formation,MTT,wound healing,and transwell assays were used to detect the proliferation,migration and invasion abilities of PC3 cells.Finally,the targeting relationship between miR-3064-5p and eukaryotic translation initiation factor5(EIF5)were confirmed by dual luciferase gene reporter assay,qRT-PCR and Western blot assays.Results Compared with normal prostate epithelial cells,the expression of miR-3064-5p in prostate cancer cells was significantly reduced(P<0.01),overexpression of miR-3064-5p could significantly inhibit the proliferation,migration and invasion of PC3 cells(P all<0.05).Bioinformatic analysis showed that EIF5 was a potential target of miR-3064-5p,and this was confirmed by dual luciferase report assay.Furthermore,overexpression of miR-3064-5p could significantly reduce protein expressions of PCNA,mmp2 and EIF5.Conclusion The expression of miR-3064-5p is downregulated in prostate cancer cells,the overexpression of miR-3064-5p may inhibit the proliferation,migration and invasion of prostate cancer through targeting EIF5 gene.