circPTPRA靶向miR-140-5p调控类风湿关节炎滑膜成纤维细胞增殖及迁移的机制研究

Mechanism of circPTPRA on regulating the proliferation and migration of rheumatoid arthritis fibroblast-like synoviocytes by targeting miR-140-5p

目的:探讨circPTPRA对类风湿关节炎滑膜成纤维细胞增殖、迁移的影响及其对miR-140-5p的调控作用。方法:原代分离培养人正常滑膜成纤维细胞与类风湿关节炎滑膜成纤维细胞,将类风湿关节炎滑膜成纤维细胞分为NC组(不转染)、si-NC组(转染si-NC)、si-circPTPRA组(转染si-circPTPRA)、miR-NC组(转染miR-NC)、miR-140-5p组(转染miR-140-5p mimics)、anti-miR-NC+si-circPTPRA组(共转染anti-miR-NC和si-circPTPRA)、anti-miR-140-5p+si-circPTPRA组(共转染anti-miR-140-5p和si-circPTPRA)。qRT-PCR检测circPTPRA与miR-140-5p的表达量;MTT实验检测细胞增殖;Transwell小室实验检测细胞迁移;双荧光素酶报告基因实验检测circPTPRA与miR-140-5p的靶向关系。结果:与正常滑膜成纤维细胞比较,类风湿关节炎滑膜成纤维细胞中circPTPRA的表达量升高,miR-140-5p的表达量降低(P<0.01);与si-NC组比较,si-circPTPRA组细胞活力降低,迁移细胞数减少(P<0.05);与miR-NC组比较,miR-140-5p组细胞活力降低,迁移细胞数减少(P<0.01);circPTPRA可靶向调控miR-140-5p;与anti-miR-NC+si-circPTPRA组比较,anti-miR-140-5p+si-circPTPRA组细胞活力升高,迁移细胞数增多(P<0.01)。结论:干扰circPTPRA表达可通过靶向调控miR-140-5p的表达而抑制类风湿关节炎滑膜成纤维细胞增殖及迁移。

Objective:To explore the effect of circPTPRA on the proliferation and migration of rheumatoid arthritis fibroblast-like synoviocytes and its regulatory effect on miR-140-5p.Methods:The primary human normal fibroblast-like synoviocyte and rheumatoid arthritis fibroblast-like synoviocyte were isolated and cultured.The synovial fibroblasts of rheumatoid arthritis were divided into NC group(without transfection),si-NC group(transfected si-NC),si-circPTPRA group(transfected si-circPTPRA),miR-NC group(transfected miR-NC)and miR-140-5p group(transfected miR-140-5p mimics),anti-miR-NC+si-circPTPRA group(co-transfected anti-miR-NC and si-circPTPRA),anti-miR-140-5p+si-circPTPRA group(co-transfected anti-miR-140-5p and si-circPTPRA).The expression levels of circPTPRA and miR-140-5p were detected by qRT-PCR.The cell proliferation was detected by MTT test,and the cell migration was tested by Transwell chamber.The targeting relationship between circPTPRA and miR-140-5p was detected through the dual luciferase reporter gene experiment.Results:Compared with normal fibroblast-like synoviocytes,in rheumatoid arthritis fibroblast-like synoviocytes,the expression of circPTPRA was increased,while the expression of miR-140-5p was decreased(P<0.01).Compared with the si-NC group,in the si-circPTPRA group,the cell viability was decreased,the number of migrating cells was decreased(P<0.05).Compared with the miR-NC group,in the miR-140-5p group,the cell viability and the number of migrating cells were decreased(P<0.01).CircPTPRA could target miR-140-5p.Compared with the anti-miR-NC+si-circPTPRA group,the cell viability was increased as well as the number of migrating cells in the anti-miR-140-5p+si-circPTPRA group(P<0.01).Conclusions:Interference with the expression of circPTPRA can inhibit the proliferation and migration of rheumatoid arthritis fibroblast-like synoviocyte by targeting the expression of miR-140-5p.