CAJ | 학술논문

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。
목적：연구microRNA-449a（miR-449a）대방광암세포J82천이능력적영향이급대파기인Notch1표체적영향。방법통과mimics전염방광암세포주J82사기과표체miR-449a，이용Transwell실험、세포화흔실험관찰세포천이능력적변화；채용실시정량PCR화단백인적검측세포Notch1표체수평적변화，병통과형광소매실험험증 miR-449a 여Notch1기인적직접조공관계。결과여대조조상비，전염miR-449a조적J82세포적천이능력감약。과표체miR-449a후， J82세포Notch1적mRNA표체무명현변화（P＝0．5739），단Notch1단백표체하조（P＝0．0135）。형광소매보고기인실험현시，miR-449a능명현억제Notch1-3’UTR적형광소매활성（P＝0．0016）。결론과표체miR-449a가능통과파향강저Notch1기인적단백표체，억제방광암세포적천이。
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.