Zhao Changfu, Wang Yang, Jin Hui, Yu Tao
Department of Orthopedics, China-Japan Union Hospital of Jilin University, Changchun 130033, China.
Department of Orthopedics, The Second Hospital of Jilin University, Changchun 130041, China.
Exp Cell Res. 2017 Oct 1;359(1):171-178. doi: 10.1016/j.yexcr.2017.07.034. Epub 2017 Jul 29.
Several microRNAs (miRs) are associated with osteoarthritis (OA) and are also functionally implicated in the pathogenesis of the disease. This study was aimed to investigate the potential roles of miR-203 in the development and progression of OA, as well as to illustrate the possible molecular mechanism. OA was simulated in human cartilage C28/I2 cells with treatments of lipopolysaccharide (LPS). LPS-induced cell injury was evaluated according to cell viability, apoptosis and release of pro-inflammatory cytokines. Expression of miR-203 after LPS treatment was assessed. Then, miR-203 was aberrantly expressed, followed by evaluation of LPS-induced cell injury. The target gene of miR-203 was virtually screened by bioinformatics method and verified by luciferase assay. Moreover, the potential associated signaling pathways were also investigated. Results showed that LPS induced decrease of cell viability and increases of cell apoptosis, release of inflammatory cytokines and expression of miR-203. LPS-induced alterations were aggravated by miR-203 overexpression but were alleviated by miR-203 inhibition. Myeloid cell leukemia-1 (MCL-1) was hypothesized and subsequently verified to be a target of miR-203, and miR-203 inhibition affected C28/I2 cells through up-regulating MCL-1. In addition, LPS-induced down-regulations of key kinases as well as phosphorylated kinases were further down-regulated by miR-203 overexpression but were abrogated by miR-203 inhibition. In conclusion, this study suggests that miR-203 suppression may inhibit the progression of OA by targeting MCL-1 and activating the Wnt/β-Catenin and JAK/STAT signal pathways.
几种微小RNA（miR）与骨关节炎（OA）相关，并且在该疾病的发病机制中也具有功能牵连。本研究旨在探讨miR-203在OA发生发展中的潜在作用，并阐明其可能的分子机制。用脂多糖（LPS）处理人软骨C28/I2细胞以模拟OA。根据细胞活力、凋亡和促炎细胞因子的释放评估LPS诱导的细胞损伤。评估LPS处理后miR-203的表达。然后，使miR-203异常表达，随后评估LPS诱导的细胞损伤。通过生物信息学方法虚拟筛选miR-203的靶基因，并通过荧光素酶测定进行验证。此外，还研究了潜在的相关信号通路。结果表明，LPS诱导细胞活力降低、细胞凋亡增加、炎性细胞因子释放增加以及miR-203表达增加。miR-203过表达加剧了LPS诱导的改变，但miR-203抑制则减轻了这些改变。推测髓样细胞白血病-1（MCL-1）并随后验证其为miR-203的靶标，并且miR-203抑制通过上调MCL-1影响C28/I2细胞。此外，miR-203过表达进一步下调了LPS诱导的关键激酶以及磷酸化激酶的表达，但miR-203抑制则消除了这种下调。总之，本研究表明，抑制miR-203可能通过靶向MCL-1并激活Wnt/β-连环蛋白和JAK/STAT信号通路来抑制OA的进展。
