Zhang Chao, Yao Zhiyong, Zhu Mingyang, Ma Xin, Shi Taoping, Li Hongzhao, Wang Baojun, Ouyang Jinzhi, Zhang Xu
Department of Urology, Chinese PLA General Hospital, Beijing, 100853, China.
Nankai University, Tianjin, 300071, China.
J Huazhong Univ Sci Technolog Med Sci. 2012 Jun;32(3):375-382. doi: 10.1007/s11596-012-0065-z. Epub 2012 Jun 9.
MicroRNAs (miRNAs or miRs) are a class of short, non-coding RNAs that participate in various oncological processes. This study aims to explore the roles of microRNA-34a (miR-34a) in invasive urothelial bladder carcinoma. miR-34a was transfected into bladder cancer cell lines 253J and J82. The miR-34a expression levels in tissues and cells were detected by using qRT-PCR. The Notch1 expression was detected by qRT-PCR and Western blotting. Cell migratory and invasive abilities were measured by Transwell chamber assay. Bioinformatics and luciferase assay were performed to predict and analyze the binding sites between miRNA-34a and Notch1. It was found that there was aberrant expression of miR-34a in bladder cancer tissues. Moreover, we revealed that ectopic expression of miR-34a suppressed cell migration and invasion, while forced expression of Notch1 increased cell migratory and invasive abilities. Finally, we observed that miR-34a transfection significantly down-regulated luciferase activity and reduced the mRNA and protein levels of Notch1. Our study concluded that microRNA-34a antagonizes Notch1 and inhibits cell migration and invasion of bladder cancer cells, which indicates the tumor-suppressive function of microRNA-34a in bladder cancer.
微小RNA(miRNA或miR)是一类短链非编码RNA,参与多种肿瘤发生过程。本研究旨在探讨微小RNA - 34a(miR - 34a)在浸润性膀胱尿路上皮癌中的作用。将miR - 34a转染至膀胱癌细胞系253J和J82。采用qRT - PCR检测组织和细胞中miR - 34a的表达水平。通过qRT - PCR和蛋白质免疫印迹法检测Notch1的表达。采用Transwell小室实验检测细胞的迁移和侵袭能力。进行生物信息学分析和荧光素酶实验以预测和分析miRNA - 34a与Notch1之间的结合位点。研究发现,miR - 34a在膀胱癌组织中存在异常表达。此外,我们发现miR - 34a的异位表达抑制细胞迁移和侵袭,而Notch1的过表达则增强细胞的迁移和侵袭能力。最后,我们观察到miR - 34a转染显著下调荧光素酶活性,并降低Notch1的mRNA和蛋白水平。我们的研究得出结论,微小RNA - 34a拮抗Notch1并抑制膀胱癌细胞的迁移和侵袭,这表明微小RNA - 34a在膀胱癌中具有肿瘤抑制功能。