Su Zhengming, Chen Duqun, Zhang Enpu, Li Yifan, Yu Zuhu, Shi Min, Jiang Zhimao, Ni Liangchao, Yang Shangqi, Gui Yaoting, Ye Jiongxian, Lai Yongqing
Department of Urology, Peking University Shenzhen Hospital, Shenzhen, Guangdong 518036, P.R. China.
Mol Med Rep. 2015 Jul;12(1):1535-43. doi: 10.3892/mmr.2015.3498. Epub 2015 Mar 17.
microRNAs (miRNAs; miR) are a class of small non-coding RNA molecules, which are involved in the pathogenesis of human diseases through the negative regulation of gene expression. Previous studies have demonstrated that miR-509-3p is a novel miRNA associated with cell proliferation and migration in 786-O renal cell carcinoma (RCC) cells. However, the mechanism of action of miR-509-3p in RCC remains to be elucidated. The present study aimed to examine the functional role and mechanism of miR-509-3p in the development of RCC. The results demonstrated that the expression levels of miR-509-3p were downregulated in the 786-O and ACHN RCC cell lines compared with the normal tissues of 10 patients with RCC, as determined by reverse transcription-quantitative polymerase chain reaction. The mRNA expression levels of mitogen-activated protein kinase kinase kinase 8 (MAP3K8) were upregulated in the RCC cell lines. Functional investigations demonstrated that the overexpression of miR-509-3p inhibited the migration and proliferation of the RCC cells, as determined by wound scratch and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays. Luciferase reporter assays revealed that the overexpression of miR-509-3p reduced the transcriptional activity of MAP3K8. Furthermore, the present study demonstrated that the ectopic transfection of miR-509-3p led to a significant reduction in the mRNA and protein expression levels of MAP3K8 in the RCC cells. Finally, knockdown of MAP3K8 inhibited the migration and proliferation of the RCC cells. Therefore, the results of the present study demonstrated that the miR-509-3p RCC suppressor was a significant regulator of the MAP3K8 oncogene, suggesting that it may have a potential therapeutic role in the treatment of RCC.
微小RNA(miRNA;miR)是一类小的非编码RNA分子,其通过对基因表达的负调控参与人类疾病的发病机制。先前的研究表明,miR-509-3p是一种与786-O肾细胞癌(RCC)细胞的增殖和迁移相关的新型miRNA。然而,miR-509-3p在RCC中的作用机制仍有待阐明。本研究旨在探讨miR-509-3p在RCC发生发展中的功能作用及机制。结果表明,通过逆转录-定量聚合酶链反应测定,与10例RCC患者的正常组织相比,786-O和ACHN RCC细胞系中miR-509-3p的表达水平下调。丝裂原活化蛋白激酶激酶激酶8(MAP3K8)的mRNA表达水平在RCC细胞系中上调。功能研究表明,通过伤口划痕和3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐试验测定,miR-509-3p的过表达抑制了RCC细胞的迁移和增殖。荧光素酶报告基因试验显示,miR-509-3p的过表达降低了MAP3K8的转录活性。此外,本研究表明,miR-509-3p的异位转染导致RCC细胞中MAP3K8的mRNA和蛋白表达水平显著降低。最后,MAP3K8的敲低抑制了RCC细胞的迁移和增殖。因此,本研究结果表明,miR-509-3p是RCC的抑制因子,是MAP3K8癌基因的重要调节因子,提示其在RCC治疗中可能具有潜在的治疗作用。