LncRNA SNHG12通过调控E2F5对前列腺癌细胞增殖、迁移和侵袭的影响

Effects of IncRNA SNHG12 on the proliferation,migration and invasiveness of prostate cancer cells by regulating E2 F5 expression

目的:分析LncRNA SNHG12通过调控E2F5对前列腺癌细胞增殖、迁移和侵袭的影响。方法:构建抑制lncRNASNHG12表达的PC3细胞,分RWPE-1组、DU145组、LNCaP组、PC3组,采用实时荧光RT-PCR检测lncRNA SNHG12、E2F5表达。并干扰E2F5表达从而对PC3细胞转染,进一步分为NC组、si-NC组、si-SNHG12组、si-E2F5组、si-SNHG12+OE-si-NC、si-SNHG12+OE-E2F5组,检测各组细胞增殖、凋亡、迁移、侵袭情况。结果:与癌旁组织相比,前列腺癌组织中lncRNA SNHG12、E2F5表达水平升高(P<0.05)。与RWPE-1组相比,DU145组、LNCaP组、PC3组细胞中LncRNA SNHG12、E2F5表达水平升高,且PC3组LncRNA SNHG12、E2F5表达水平最高(P<0.05)。与si-NC组相比,si-SNHG12组SNHG12表达水平降低(P<0.05),表明干扰lncRNA SNHG12表达的PC3细胞株构建成功,与si-NC组相比,si-SNHG12组CyclinD1、MMP-9、OD值、迁移细胞数、侵袭细胞数降低,凋亡细胞升高(P<0.05)。与si-NC组相比,si-E2F5组E2F5表达水平降低(P<0.05),表明干扰E2F5的PC3细胞株构建成功,与si-NC组相比,si-E2F5组CyclinD1、MMP-9、OD值、迁移细胞数、侵袭细胞数降低,凋亡细胞升高(P<0.05)。双荧光素酶报告试验显示,与si-NC组相比,E2F5可使SNHG12荧光素酶活性降低(P<0.05),对MUT-SNHG12荧光素酶活性影响较小(P>0.05)。与si-NC组相比,抑制lncRNA SNHG12表达可使PC3细胞中E2F5表达、CyclinD1、MMP-9、OD值、迁移细胞数、侵袭细胞数降低,凋亡细胞升高(P<0.05),与si-SNHG12+OE-si-NC组相比,si-SNHG12+OE-E2F5组E2F5表达、CyclinD1、MMP-9、OD值、迁移细胞数、侵袭细胞数升高,凋亡细胞降低(P<0.05)。结论:干扰lncRNA SNHG12表达,可对E2F5表达进行调控,抑制前列腺癌细胞增殖、迁移、侵袭,并促进其癌细胞的凋亡。

Objective:To analyze the effects of IncRNA SNHG12 on the proliferation,migration and invasiveness of PCa cells by regulating the expression of E2F5.Methods:Using real time fluorescence RT-PCR,we detected the expressions of IncRNA SNHG12 and E2F5,constructed the PC3 cells inhibiting the lncRNA SNHG12 expression.After transfection of the PC3 cells,we divided them into an NC,a si-NC,a si-SNHG12,a si-E2F5,a si-SNHG12+OE-si-NC,and a si-SNHG12+OE-E2F5 group,followed by examination of the proliferation,apoptosis,migration and invasiveness of the cells in different groups.Results:Theexpressions of IncRNA SNHG12 and E2F5 were significantly up-regulated in the PCa tissue compared with those in the adjacent tissue(P<0.05),remarkably higher in the DU145,LNCaP and PC3 groups than in the RWPE-1 group,the highest in the PC3 group(P<0.05).The expression of SNHG12 was markedly down-regulated in the si-SNHC12 group(P<0.05)in comparison with that in the si-NC group,indicating the successful construction of a PC3 cell line interfering with the lncRNA SNHG12 expression.Compared with the si-NC group,the si-SNHG12 group showed significant decreases in the values of CyclinD1,MMP-9 and OD and the numbers of migrating and invading cells,and an increase in apoptotic cells(P<0.05),while the si-E2F5 group exhibited a remarkably down-regulated expression of E2F5(P<0.05),reduced values of CyclinD1,MMP-9 and OD,decreased numbers of migrating and invading cells and an increased number of apoptotic cells(P<0.05).The dual luciferase report test showed that E2F5 reduced the luciferase activity of SNHG12(P<0.05 and had an insignificant impact on the luciferase activity of MUT-SNHG12(P>0.05).Inhibiting the expression of lncRNA SNHG12 resulted in significant decreases in the expression of E2F5,values of CyclinDI,MMP-9 and OD and numbers of migrating and invading cells,but an increase in apoptotic cells(P<0.05).The E2F5 expression,the CyclinD1,MMP-9 and OD values and the numbers of migrating and invading cells were markedly increased while the number of apoptotic cells decreased in the si-SNHC12+OE-E2F5 group compared with those in the si-SNHG12+OE-si-NC group(P<0.05).Conclusion:Interfering with the expression of IncRNA SNHG12 can regulate that of E2F5,inhibit the proliferation,migration and invasiveness of PCa cells and promote their apoptosis.