Feng Lei, Jing Wenseng, Jin Shengyu, Wang Bo
Department of Joint Surgery, Honghui Hospital, Xi'an Jiaotong University, No. 555 Youyi East Road, Xi'an, Shaanxi, 710054, China.
Inflammation. 2023 Feb;46(1):161-174. doi: 10.1007/s10753-022-01719-9. Epub 2022 Aug 2.
Dysregulation of circular RNAs (circRNAs) has been observed in multiple diseases including rheumatoid arthritis (RA), and we investigated the role of the circ_0088194/microRNA (miR)-30a-3p/a disintegrin and metalloproteinase 10 (ADAM10) axis in RA. Circ_0088194, miR-30a-3p, and ADAM10 contents in RA tissues and RA-fibroblast-like synoviocytes (RA-FLSs) were analyzed by real-time quantitative polymerase chain reaction (RT-qPCR) and western blot. Cell proliferation, migration, apoptosis, and inflammatory factor secretion of RA-FLSs were detected using 5-ethynyl-2'-deoxyuridine (EdU), wound healing assay, flow cytometry, and enzyme-linked immunosorbent assay (ELISA). Targeting relationship between miR-30a-3p and circ_0088194 or ADAM10 was validated by luciferase reporter system, RNA immunoprecipitation (RIP), and RNA pull-down assays. Circ_0088194 and ADAM10 levels were increased, while miR-30a-3p was decreased in RA tissues and RA-FLSs. Circ_0088194 knockdown suppressed the growth, migration, and inflammation of RA-FLSs, while the upregulation of circ_0088194 showed opposite effects. Circ_0088194 directly targeted miR-30a-3p, ADAM10 was a target of miR-30a-3p, and circ_0088194 regulated the expression of ADAM10 by sponging miR-30a-3p. MiR-30a-3p inhibition restored the inhibition effects of circ_0088194 knockdown or RA-FLSs. Moreover, miR-30a-3p re-expression repressed growth, migration, and inflammatory response in RA-FLSs, which were reversed by ADAM10 overexpression. Circ_0088194 acted on miR-30a-3p/ADAM10 axis to promote the proliferation, migration, and inflammatory response, and inhibit apoptosis in RA-FLSs.
在包括类风湿性关节炎（RA）在内的多种疾病中均观察到环状RNA（circRNA）失调，我们研究了circ_0088194/微小RNA（miR）-30a-3p/解聚素和金属蛋白酶10（ADAM10）轴在类风湿性关节炎中的作用。通过实时定量聚合酶链反应（RT-qPCR）和蛋白质印迹法分析类风湿性关节炎组织和类风湿性成纤维样滑膜细胞（RA-FLSs）中circ_0088194、miR-30a-3p和ADAM10的含量。使用5-乙炔基-2'-脱氧尿苷（EdU）、伤口愈合试验、流式细胞术和酶联免疫吸附测定（ELISA）检测RA-FLSs的细胞增殖、迁移、凋亡和炎症因子分泌。通过荧光素酶报告系统、RNA免疫沉淀（RIP）和RNA下拉试验验证miR-30a-3p与circ_0088194或ADAM10之间的靶向关系。在类风湿性关节炎组织和RA-FLSs中，circ_0088194和ADAM10水平升高，而miR-30a-3p水平降低。敲低circ_0088194可抑制RA-FLSs的生长、迁移和炎症，而circ_0088194的上调则显示出相反的效果。circ_0088194直接靶向miR-30a-3p，ADAM10是miR-30a-3p的靶标，circ_0088194通过海绵吸附miR-30a-3p来调节ADAM10的表达。抑制miR-30a-3p可恢复敲低circ_0088194或RA-FLSs的抑制作用。此外，miR-30a-3p的重新表达可抑制RA-FLSs的生长、迁移和炎症反应，而ADAM10的过表达可逆转这些作用。circ_0088194作用于miR-30a-3p/ADAM轴，以促进RA-FLSs的增殖、迁移和炎症反应，并抑制其凋亡。
