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目的:探讨miR-519d对膀胱癌细胞系5637增殖的影响及分子机制。方法:通过转染miRNA mimics在膀胱癌细胞系5637中过表达miR-519d,分别利用MTS及细胞集落形成试验,检测膀胱癌细胞系5637增殖能力的改变。通过双荧光素酶报道实验和Western Blot方法验证miR-519d对OCT4的靶向调控。利用OCT4特异性干扰RNA证实miR-519d通过靶向调控OCT4抑制膀胱癌细胞系5673增殖。结果:在膀胱癌细胞系过表达miR-519d后,细胞活性、增殖能力明显下降;双荧光素酶报道实验结果表明,相对于各对照组,共转染OCT4 3'UTR载体与miR-519d组荧光素酶相对活性明显降低(P<0.05)。过表达miR-519d后5637细胞中OCT4蛋白表达下调。利用siRNA特异性下调OCT4表达后,5637细胞活性及增殖能力同样受到抑制。结论:miR-519d能靶向结合OCT4 3'UTR序列,通过下调OCT4的表达抑制膀胱癌细胞系5637的增殖。
Objective: To investigate the effect of miR-519d on the proliferation of bladder cancer cell line 5637 and its molecular mechanism. Methods: miR-519d was overexpressed in transfected bladder cancer cell line 5637 by transfection of miRNA mimics. MTS and colony formation assay were used to detect the proliferation of bladder cancer cell line 5637. Targeting regulation of OCT4 by miR-519d was verified by dual luciferase reporter assay and Western Blot assay. Using OCT4-specific interfering RNAs confirmed that miR-519d inhibits the proliferation of bladder cancer cell line 5673 by targeting OCT4. Results: The proliferation and cell viability decreased after miR-519d overexpression in bladder cancer cell lines. The results of dual luciferase reporter assay showed that OCT4 3'UTR vector and miR-519d co-transfected with each control group The relative luciferase activity was significantly decreased (P <0.05). The OCT4 protein expression was down-regulated in 5637 cells after overexpression of miR-519d. Specific siRNA down-regulation OCT4 expression, 5637 cell activity and proliferation were also inhibited. Conclusion: miR-519d can bind to OCT4 3'UTR sequence and inhibit the proliferation of bladder cancer cell line 5637 by down-regulating OCT4 expression.