摘要
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 mechanism of suppression of miR-519d in bladder cancer cell line 5637. Method： After transfected with miR-519d mimics in bladder cancer cell line 5637, the proliferation of 5637 cells were examined by MTS assay and colony formation experiment. Western blot assay was used to analyze the expression of OCT4 in transfected cells, and the relative luciferase activity was detected by dual luciferase reporter system. OCT4-specific siRNA was used as a positive control. Result： The proliferation and colony formation po- tential of 5637 cells transfected with miR-519d mimics were significantly lower than control group. The relative lu- ciferase activities of OCT4-3UTR/miR-519d co-transfected group were significantly lower than other groups （P〈 0.05）, and the expression of OCT4 has been down-regulated by miR-519d transfection. And the effects of OCT4- specific siRNA on cell proliferation and colony formation complied with that of miR-519d. Conclusion： It is indica- ted that miR-519d suppresses cell growth of bladder cancer cell 5637 directly through targeting OCT4.
出处
《临床泌尿外科杂志》
2014年第2期163-166,共4页
Journal of Clinical Urology
