Yang Yan, Wang Yanfeng, Liang Qingwei, Yao Lutian, Gu Shizhong, Bai Xizhuang
Department of Sports Medicine and Joint Surgery/Orthopaedics, First Affiliated Hospital of China Medical University, Shenyang, Liaoning, 110001, China.
Department of Sports Medicine/Joint Surgery, The People's Hospital of China Medical University, Shenyang, Liaoning, 110001, China.
J Cell Biochem. 2017 Aug;118(8):2295-2301. doi: 10.1002/jcb.25883. Epub 2017 Mar 3.
Our purpose is to study the roles of microRNA-338-5p (miR-338-5p) on the proliferation, invasion, and inflammatory response of fibroblast-like synoviocytes (SFs) in rheumatoid arthritis patients by regulating SPRY1. The target relationship between miR-338-5p and SPRY1 was validated through luciferase reporter system. The expression of miR-338-5p and SPRY1 in synovial tissues and synovial cells were detected using RT-PCR and western blot. The mimics and inhibitors of miR-338-5p were transfected into SFs. MTT, Transwell, and ELISA assays were used to analyze cell proliferation, invasiveness, and the secreted extracellular pro-inflammatory cytokines (such as IL-1a, IL-6, COX2) levels of SFs. MiR-338-5p was highly expressed in rheumatoid arthritis tissues and cells, and directly down-regulated the expression of SPRY1 in the SFs of rheumatoid arthritis patients. Cell proliferation, invasiveness and the expression level of pro-inflammatory cytokines in synovial cells increased after the transfection of miR-338-5p mimics, while the proliferation, invasion and expression level of pro-inflammatory cytokines decreased after the transfection of miR-338-5p inhibitors. In conclusion,miR-338-5p promoted the proliferation, invasion and inflammatory reaction in SFs of rheumatoid arthritis by directly down-regulating SPRY1 expression. J. Cell. Biochem. 118: 2295-2301, 2017. © 2017 Wiley Periodicals, Inc.
我们的目的是通过调节SPRY1来研究微小RNA - 338 - 5p(miR - 338 - 5p)对类风湿性关节炎患者成纤维样滑膜细胞(SFs)增殖、侵袭及炎症反应的作用。通过荧光素酶报告系统验证了miR - 338 - 5p与SPRY1之间的靶向关系。采用逆转录聚合酶链反应(RT - PCR)和蛋白质免疫印迹法检测滑膜组织及滑膜细胞中miR - 338 - 5p和SPRY1的表达。将miR - 338 - 5p的模拟物和抑制剂转染到SFs中。采用MTT法、Transwell法和酶联免疫吸附测定(ELISA)法分析SFs的细胞增殖、侵袭能力以及分泌的细胞外促炎细胞因子(如IL - 1α、IL - 6、COX2)水平。miR - 338 - 5p在类风湿性关节炎组织和细胞中高表达,并直接下调类风湿性关节炎患者SFs中SPRY1的表达。转染miR - 338 - 5p模拟物后,滑膜细胞的增殖、侵袭能力及促炎细胞因子表达水平升高,而转染miR - 338 - 5p抑制剂后,增殖、侵袭能力及促炎细胞因子表达水平降低。总之,miR - 338 - 5p通过直接下调SPRY1表达促进类风湿性关节炎SFs的增殖、侵袭及炎症反应。《细胞生物化学杂志》118: 2295 - 2301, 2017。© 2017威利期刊公司