Li Congda, Ma Deying, Yang Jinhu, Lin Xiangbo, Chen Bo
Department of Orthopedic, People's Hospital of Rizhao, Rizhao, Shandong 276800, P.R. China.
Oncol Lett. 2018 Jul;16(1):829-834. doi: 10.3892/ol.2018.8694. Epub 2018 May 10.
Many studies have shown that microRNA regulates the development and treatment of osteosarcoma (OS). In many human cancer studies, the expression of microRNA-202 has been shown to be abnormal. The aim of the study was to examine the role of miR-202-5p in the occurrence and formation of OS. miR-202-5p and Rho-associated coiled-coil containing protein kinase 1 (ROCK1) levels were assessed using RT-qPCR in OS tissues and cell lines. The cell migrating and invasive abilities were detected by the Transwell assay in OS. Moreover, the relationship between miR-202-5p and ROCK1 was verified via luciferase reporter assay. The protein level of ROCK1 was identified by western blot analysis. Downregulation of miR-202-5p was identified in OS tissues and cell lines. In addition, the miR-202-5p overexpression had inhibitory action for cell migration and invasion in OS. Moreover, miR-202-5p directly targeted ROCK1 and negatively regulated its expression. Upregulation of ROCK1 had a carcinogenic effect in OS. Furthermore, the upregulation of ROCK1 restored the suppressive effect of miR-202-5p. miR-202-5p, in turn, weakened the abilities of cell migration and invasion in OS by inhibiting ROCK1 expression. As a result, miR-202-5p may be developed as a potential pathway in the reatment of OS.
许多研究表明,微小RNA可调节骨肉瘤(OS)的发展及治疗。在许多人类癌症研究中,已表明微小RNA-202的表达存在异常。本研究的目的是检测miR-202-5p在OS发生及形成过程中的作用。采用RT-qPCR检测OS组织及细胞系中miR-202-5p和含Rho相关卷曲螺旋蛋白激酶1(ROCK1)的水平。通过Transwell实验检测OS细胞的迁移和侵袭能力。此外,通过荧光素酶报告基因实验验证miR-202-5p与ROCK1之间的关系。通过蛋白质免疫印迹分析鉴定ROCK1的蛋白水平。在OS组织及细胞系中发现miR-202-5p表达下调。此外,miR-202-5p过表达对OS细胞的迁移和侵袭具有抑制作用。而且,miR-202-5p直接靶向ROCK1并对其表达起负向调节作用。ROCK1的上调在OS中具有致癌作用。此外,ROCK1的上调恢复了miR-202-5p的抑制作用。反过来,miR-202-5p通过抑制ROCK1表达减弱了OS细胞的迁移和侵袭能力。因此,miR-202-5p可能会被开发成为治疗OS的潜在途径。