Zeng Tao, Peng Lifen, Chao Chaohai, Fu Bin, Wang Gongxian, Wang Yibing, Zhu Xinshen
School of Medicine, Nanchang University Nanjing, Jiangsu, PR China ; Department of Urology, The People's Hospital of Jiangxi Province Nanchang, PR China.
Department of E.N.T., The People's Hospital of Jiangxi Province Nanchang, PR China.
Int J Clin Exp Pathol. 2014 Oct 15;7(11):7653-62. eCollection 2014.
MicroRNAs (miRNAs) have recently been reported play a crucial role in some tumors. In order to investigate the association of miR-451 with bladder cancer, we investigate the expression of miR-451 in bladder cancer tissues and its role in biological behavior of T24, 5637 and J28 bladder cancer cell lines. Quantitative RT-PCR results showed miR-451 was significantly down-regulated in bladder cancer tissues and paracancerous tissues compared with normal bladder tissues. miR-451 expression was significantly associated with histological differentiation degree and TNM stage. Over-expression of miR-451 was established by transfecting miR-451 mimics into T24, 5637 and J28 cells, and its effects on the biological behavior of bladder cancer were studied using transwell assay, migration assay, adhesion assay, MTT and flow cytometry. Results indicated over-expression of miR-451 significantly inhibited cell proliferation, migration, invasion and induced apoptosis of the bladder cancer cells. Furthermore, we investigated the expression level of EMT related proteins in transfected 5637 cells by western blot. Results shown E-cadherin was up-regulated more significantly than N-cadherin, vimentin and Snail. N-cadherin and vimentin were up-regulated significantly when miR-451 was inhibited in miR-451 inhibitor group, however, no significant changes in mimics group. In conclusion, miR451 should be a tumor-suppressing gene in bladder cancer. miR-451 could maintain the bladder tumor cells in epithelial phenotype, inhibit EMT process, thereby reducing the invasion and migration of tumor cells.
最近有报道称,微小RNA(miRNA)在某些肿瘤中起着至关重要的作用。为了研究miR-451与膀胱癌的关系,我们研究了miR-451在膀胱癌组织中的表达及其在T24、5637和J28膀胱癌细胞系生物学行为中的作用。定量逆转录聚合酶链反应(RT-PCR)结果显示,与正常膀胱组织相比,miR-451在膀胱癌组织和癌旁组织中显著下调。miR-451表达与组织学分化程度和TNM分期显著相关。通过将miR-451模拟物转染到T24、5637和J28细胞中建立miR-451的过表达,并使用Transwell实验、迁移实验、黏附实验、MTT和流式细胞术研究其对膀胱癌细胞生物学行为的影响。结果表明,miR-451的过表达显著抑制膀胱癌细胞的增殖、迁移、侵袭并诱导其凋亡。此外,我们通过蛋白质免疫印迹法检测了转染的5637细胞中上皮-间质转化(EMT)相关蛋白的表达水平。结果显示,E-钙黏蛋白上调比N-钙黏蛋白、波形蛋白和Snail更显著。在miR-451抑制剂组中抑制miR-451时,N-钙黏蛋白和波形蛋白显著上调,但在模拟物组中无显著变化。总之,miR-451在膀胱癌中应是一种肿瘤抑制基因。miR-451可使膀胱肿瘤细胞维持上皮表型,抑制EMT过程,从而减少肿瘤细胞的侵袭和迁移。