微RNA-139-5p通过靶向ITGA3抑制膀胱癌细胞增殖、迁移和侵袭的研究机制

MicroRNA-139-5p inhibits proliferation,migration and invasion of bladder cancer cells by targeting ITGA3:a mechanistic study

目的探讨微RNA(miR)-139-5p对膀胱癌细胞增殖、迁移和侵袭的作用机制.方法运用qRT-PCR法检测膀胱癌细胞、膀胱上皮细胞中miR-139-5p的表达;将miR-139-5p组(转染miR-139-5p mimics)、miR-NC组(转染miR-NC)、si-con组(转染si-con)、si-ITGA3组(转染si-ITGA3)、miR-139-5p+pcDNA组(共转染miR-139-5p mimics和pcDNA)、miR-139-5p+pcDNA-ITGA3组(共转染miR-139-5p mimics和pcDNA-ITGA3),用脂质体法转染至T24细胞;免疫印迹检测细胞中ITGA3的蛋白表达;MTT法检测细胞增殖;Transwell法检测细胞迁移和侵袭;双荧光素酶报告基因检测实验检测细胞的荧光活性.结果与膀胱上皮细胞相比,膀胱癌细胞中miR-139-5p表达明显降低,ITGA3表达明显升高(P<0.05);过表达miR-139-5p、敲减ITGA3均可抑制T24细胞的增殖、迁移和侵袭;miR-139-5p可抑制野生型ITGA3细胞的荧光活性,并且负向调控ITGA3的表达;过表达ITGA3可逆转miR-139-5p对T24细胞增殖、迁移、侵袭的抑制作用.结论miR-139-5p可抑制膀胱癌细胞的增殖、迁移、侵袭,其机制与靶向负调控ITGA3有关,将可为膀胱癌的诊断和治疗提供靶点.

Objective To investigate the mechanism underlying the effects of miR-139-5p on proliferation,migration and invasion of bladder cancer cells.Methods The expression of miR-139-5p in bladder cancer cells and bladder epithelial cells was detected by qRT-PCR.Liposome transfection of T24 cells led to assignment of several study groups:the miR-139-5p group(transfected with miR-139-5p mimics),the miR-NC group(transfected with miR-NC),the si-con group(transfected with si-con),the si-ITGA3 group(transfected with si-ITGA3),the miR-139-5p+pcDNA group(co-transfected with miR-139-5p mimics and pcDNA),and the miR-139-5p+pcDNA-ITGA3 group(co-transfected with miR-139-5p mimics and pcDNA-ITGA3).Western blot was used to detect the expression of ITGA3 protein in T24 cells;MTT assay was used to detect cell proliferation;Transwell method was used to detect cell migration and invasion;dual luciferase reporter assay was used to detect the fluorescence activity of cells.Results Compared with bladder epithelial cells,the expression of miR-139-5p was significantly decreased and the expression of ITGA3 was significantly increased in bladder cancer cells(P<0.05).miR-139-5p overexpression or ITGA3 knockdown inhibited proliferation,migration and invasion of T24 cells.miR-139-5p may inhibit the fluorescence activity of wild-type ITGA3 cells and negatively regulate the expression of ITGA3.ITGA3 overexpression may reverse the inhibitory effects of miR-139-5p on proliferation,migration and invasion of T24 cells.Conclusion miR-139-5p may inhibit the proliferation,migration and invasion of bladder cancer cells.The mechanism is related to the negative targeted regulation of ITGA3,which will provide a target for the diagnosis and treatment of bladder cancer.