Objective To investigate the effect of miR-183 on the invasion of prostate cancer cells and study its target gene EPHA4 and its function.Methods The expression levels of miR-183 and EPHA4 in elderly prostate cancer and paracancerous tissues were detected by real-time fluorescence quantitative PCR. PC3 and LNCaP were elderly prostate cancer cell lines that were capable of transfecting miR-183ASO,cell motility and invasiveness were evaluated by negative control transwell migration and invasion assays. Then real-time fluores-cence quantitative PCR,Western blot and GFP reporter were used to verify the regulation of miRHA-183 on its target gene EPHA4. Results Real-time quantitative PCR showed that miR-183 was highly expressed in elderly prostate cancer (P<0.01). Migration and invasion of miR-183 ASO cells were decreased significantly compared with the control group(P<0.01). Bioinformatics revealed that the EPHA4 3'UTR contained a potential binding site for miR-183. The fluorescence intensity of EPHA4 3'UTR in miR-183 ASO transfected cells were decreased significantly compared with the control group(P<0.01),miR-183 minmcs were increased the fluorescence intensity of EPHA4 3'UTR(P<0.01),While there was no significant effect on the mutant 3'UTR. EPHA4 mRNA level was decreased in miR-183 ASO,while increased in miR-183 mimics. The protein level of EPHA4 in miR-183 ASO group was lower than that in control group(P<0.01). Conclusions miR-183 regulates the migration and invasion of elderly prostate cancer cells through EPHA4.%目的 探讨微小RNA(miR)-183对老年前列腺癌细胞转移中的影响及其与靶基因促红细胞生成素产生肝细胞受体(EPH)A4的价值.方法 对老年前列腺癌和癌旁组织中miR-183和EPHA4的表达水平通过实时荧光定量PCR(RT-PCR)方法进行检测.老年前列腺癌细胞系PC3和LNCaP能够转染miR-183抗链球菌溶血素(AS)O,通过与阴性对照及Transwell迁移和侵袭实验对其细胞的运动和侵袭能力进行评价.再通过使用RT-PCR、Western印迹、绿色荧光蛋白(GFP)报告载体3种实验验证miR-183对其靶基因EPHA4的调控.结果 RT-PCR显示miR-183处于高表达水平(P<0.01).转染miR-183 ASO细胞的迁移和侵袭能力显著低于对照组(P<0.01).生物信息学显示EPHA43′UTR含有miR-183的潜在结合位点.转染 miR-183 ASO组细胞EPHA43′UTR 荧光强度显著低于对照组(P<0.01),miR-183 minmcs增加EPHA43′UTR的荧光强度(P<0.01),而对突变的3′UTR无明显影响.miR-183 ASO中EPHA4 mRNA水平降低,miR-183 mimics中EPHA4 mRNA水平升高.miR-183 ASO组EPHA4的蛋白含量较对照组显著下降(P<0.01).结论 miR-183通过EPHA4对老年前列腺癌细胞的迁移和侵袭进行调控,发挥着癌基因的作用.
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