miR-6883-5p在膀胱癌患者血清中的表达及其对膀胱癌细胞迁移和增殖的影响

The expression of miR-6883-5p in the serum of bladder cancer patients and its effect on the migration and proliferation of bladder cancer cells

目的探究微小RNA(miRNA,miR)-6883-5p在膀胱癌患者血清中的表达及其对膀胱癌细胞迁移和增殖能力的影响。方法利用实时荧光定量PCR(qRT-PCR)法检测miR-6883-5p在39例膀胱癌患者和39例同期健康体检者血清中的表达水平。利用qRT-PCR法检测miR-6883-5p在正常膀胱黏膜上皮细胞和膀胱癌细胞中的表达水平。向miR-6883-5p表达最低的膀胱癌细胞转染miR-6883-5p mimics或阴性对照(NC),即miR-6883-5p组和NC组。利用Transwell迁移实验和CCK-8比色法检测转染miR-6883-5p后细胞的迁移和增殖能力。利用生物信息学软件和双荧光素酶报告基因预测和检验miR-6883-5p的靶基因。利用qRT-PCR和Western blot分别检测转染miR-6883-5p后细胞中靶基因的表达水平。结果膀胱癌患者血清中miR-6883-5p的表达水平显著低于健康体检者(P<0.01)。膀胱癌细胞中miR-6883-5p的表达水平均显著低于正常膀胱黏膜上皮细胞(均P<0.05),miR-6883-5p表达水平最低的细胞是5637细胞(P<0.01)。转染miR-6883-5p后,miR-6883-5p组细胞的迁移和增殖能力均显著低于NC组(均P<0.05)。磷脂酰基醇蛋白聚糖-6(GPC6)是miR-6883-5p的靶基因。转染miR-6883-5p后,miR-6883-5p组细胞中GPC6 mRNA和蛋白的表达水平均显著低于NC组(均P<0.01)。结论miR-6883-5p在膀胱癌患者血清中显著低表达,miR-6883-5p与膀胱癌的发生发展相关;过表达miR-6883-5p可以抑制膀胱癌细胞的迁移和增殖;miR-6883-5p可能通过下调GPC6的表达抑制膀胱癌的发生发展。

Objective To explore the expression of miR-6883-5p in the serum of bladder cancer patients and its effect on the migration and proliferation ability of bladder cancer cells.Methods The real-time fluorescent quantitative polymerase chain reaction(qRT-PCR)method was used to detect the expression level of miR-6883-5p in the serum of 39 patients with bladder cancer and 39 healthy subjects,as well as the expression level of miR-6883-5p in normal bladder mucosal epithelial cells and bladder cancer cells.The bladder cancer cells with the lowest expression of miR-6883-5p were transfected with miR-6883-5p mimics or negative control(NC),namely miR-6883-5p group and NC group.Transwell migration experiment and cell counting kit-8(CCK-8)colorimetric method were used to detect the migration and proliferation ability of cells transfected with miR-6883-5p.Bioinformatics software and dual luciferase reporter gene were used to predict and test the target genes of miR-6883-5p.qRT-PCR and Western blot were used to detect the expression levels of target genes in cells transfected with miR-6883-5p.Results The expression level of miR-6883-5p in the serum of bladder cancer patients was significantly lower than that of healthy persons(P<0.01).The expression level of miR-6883-5p in bladder cancer cells was significantly lower than that in normal bladder mucosal epithelial cells(all P<0.05),and the lowest expression of miR-6883-5p was in 5637 cells(P<0.01).After transfection with miR-6883-5p,the migration and proliferation of cells in miR-6883-5p group were significantly lower than those in NC group(all P<0.05).Phosphatidyl alcohol proteoglycan-6(GPC6)was the target gene of miR-6883-5p.After transfection with miR-6883-5p,the expression levels of GPC6 mRNA and protein in miR-6883-5p group significantly decreased(P<0.01).Conclusions miR-6883-5p is significantly low expressed in the serum of bladder cancer patients,and miR-6883-5p is related to the occurrence and development of bladder cancer;overexpression of miR-6883-5p can inhibit the migration and proliferation of bladder cancer cells;miR-6883-5p can inhibit the occurrence and development of bladder cancer by down-regulating the expression of GPC6.