Abstract:

【Abstract】〓Objective〓To explore the influence of microRNA-449a (miR-449a) on migratory ability of bladder cancer cell line J82 and evaluate the correlation between miR-449a and Notch1. Methods〓After transfection of microRNA mimics into J82 cells, cell migratory ability was assessed by transwell chamber assay and wound healing assay. Expression of Notch1 was evaluated by quantitative real-time polymerase chain reaction and western blotting. Luciferase assay was used to confirm the relationship between miR-449a and Notch1. Results〓Transwell chamber assays and wound healing assays revealed that transfection of miR-449a suppressed the cell migration of J82 cells. Ectopic overexpression of miR-449a caused significant decrease of protein levels of Notch1 (P=0.0135), while the mRNA levels of Notch1 showed little change.(P=0.5739). In addition, luciferase assays showed that miR-449a significantly reduced the normalized Notch1 3'UTR luciferase activity.(P=0.0016). Conclusion〓Overexpressed miR-449a can inhibit migration of bladder cancer cells by antagonizing Notch1,.thus indicating its tumor suppressive function in bladder cancer.

Key words: microRNA｜ Bladder cancer｜ Cell migration｜ Notch1

摘要：

【摘要】〓目的〓研究microRNA-449a(miR-449a)对膀胱癌细胞J82迁移能力的影响以及对靶基因Notch1表达的影响。方法〓通过mimics转染膀胱癌细胞株J82使其过表达miR-449a,利用Transwell实验、细胞划痕实验观察细胞迁移能力的变化;采用实时定量PCR和蛋白印迹检测细胞Notch1表达水平的变化,并通过荧光素酶实验验证miR-449a与Notch1基因的直接调控关系。结果〓与对照组相比,转染miR-449a组的J82细胞的迁移能力减弱。过表达miR-449a后,J82细胞Notch1的mRNA表达无明显变化(P=0.5739),但Notch1蛋白表达下调(P=0.0135)。荧光素酶报告基因实验显示,miR-449a能明显抑制Notch1-3’UTR的荧光素酶活性(P=0.0016)。结论〓过表达miR-449a可能通过靶向降低Notch1基因的蛋白表达,抑制膀胱癌细胞的迁移。

关键词: 微小RNA, 膀胱癌, 细胞迁移, Notch1