Department of Orthopaedics, Honghui Hospital, Xi'an Jiaotong University, No.555 Youyi Dong Lu, Nanshaomen, Xi'an City, Xi'an 710054, Shaanxi, China.
Department of Emergency, The First Affiliated Hospital of Xinxiang Medical University, Weihui City, 453100, Henan, China.
Biochem Genet. 2023 Apr;61(2):521-537. doi: 10.1007/s10528-022-10265-w. Epub 2022 Aug 19.
Circular RNA_0004712 (circ_0004712) is reported to be up-regulated in rheumatoid arthritis (RA) patients. Nevertheless, its role and mechanism in RA pathology remain to be clarified. RNA and protein expression was determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot assay. Cell viability, proliferation, apoptosis, migration, and inflammation were assessed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, 5-ethynyl-20-deoxyuridine assay, flow cytometry, scratch test, and enzyme-linked immunosorbent assay. The target correlation between microRNA-633 (miR-633) and circ_0004712 or TNF receptor associated factor 6 (TRAF6) was verified by dual-luciferase reporter assay and RNA immunoprecipitation assay. Circ_0004712 was up-regulated in RA synovial tissues and RA fibroblast-like synoviocytes (RA-FLSs). Circ_0004712 silencing suppressed the viability, proliferation, migration and inflammatory response and facilitated the apoptosis of RA-FLSs. miR-633 was confirmed to be a direct target of circ_0004712, and miR-633 knockdown reversed circ_0004712 silencing-mediated protective effects on the dysfunction and inflammation of RA-FLSs. TRAF6 was a direct target of miR-633, and miR-633 overexpression suppressed the aggressive changes of RA-FLSs by down-regulating TRAF6. Circ_0004712 could up-regulate TRAF6 expression by sponging miR-633 in RA-FLSs. Circ_0004712 interference inactivated nuclear factor (NF)-κB signaling by targeting miR-633/TRAF6 axis. Circ_0004712 silencing inhibited the aggressive changes of RA-FLSs by targeting miR-633/TRAF6 axis and NF-κB signaling, which provided new targets for RA therapy.
环状 RNA_0004712(circ_0004712)在类风湿关节炎(RA)患者中呈上调表达。然而,其在 RA 病理中的作用和机制仍有待阐明。通过逆转录定量聚合酶链反应(RT-qPCR)和 Western blot 检测 RNA 和蛋白表达。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)比色法、5-乙炔基-20-脱氧尿苷(EdU)检测法、流式细胞术、划痕试验和酶联免疫吸附试验(ELISA)评估细胞活力、增殖、凋亡、迁移和炎症。通过双荧光素酶报告基因检测和 RNA 免疫沉淀试验验证 microRNA-633(miR-633)与 circ_0004712 或 TNF 受体相关因子 6(TRAF6)之间的靶相关性。Circ_0004712 在 RA 滑膜组织和 RA 成纤维样滑膜细胞(RA-FLSs)中呈上调表达。Circ_0004712 沉默抑制 RA-FLSs 的活力、增殖、迁移和炎症反应,并促进其凋亡。miR-633 被证实是 circ_0004712 的直接靶标,miR-633 敲低逆转了 circ_0004712 沉默对 RA-FLSs 功能障碍和炎症的保护作用。TRAF6 是 miR-633 的直接靶标,miR-633 过表达通过下调 TRAF6 抑制 RA-FLSs 的侵袭性变化。Circ_0004712 通过海绵吸附 miR-633 在 RA-FLSs 中上调 TRAF6 表达。Circ_0004712 干扰通过靶向 miR-633/TRAF6 轴抑制 NF-κB 信号。Circ_0004712 沉默通过靶向 miR-633/TRAF6 轴和 NF-κB 信号抑制 RA-FLSs 的侵袭性变化,为 RA 治疗提供了新的靶点。