Geng Xueli, Zhao Chunnan, Zhang Zezhi, Liu Yanling, Zhang Xiuqin, Ding Peijian
Department of Clinical Laboratory, Affiliated Hospital of Chengde Medical College, Chengde, China.
Department of Rheumatism Immunity, Affiliated Hospital of Chengde Medical College, Hebei, China.
Autoimmunity. 2022 May;55(3):157-167. doi: 10.1080/08916934.2022.2027920. Epub 2022 Mar 30.
The function and pathological significance of circular RNAs (circRNAs) in autoimmune diseases, such as rheumatoid arthritis (RA), are barely known. Here, we explored the role of circ_0088036 in RA progression and its associated mechanism.
The synovial lining layer tissues of RA patients and non-RA control patients were collected for clinical study , and tumour necrosis factor α (TNF-α)-induced RA-fibroblast-like synoviocytes (RA-FLSs) were used for the experiments . Cell proliferation was assessed by Cell Counting Kit 8 (CCK8) assay and flow cytometry. Cell apoptosis was analyzed by flow cytometry. Enzyme-linked immunosorbent assay (ELISA) was conducted to analyze the release of pro-inflammatory cytokines. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were performed to verify the target interaction between microRNA-326 (miR-326) and circ_0088036 or frizzled class receptor 4 (FZD4).
Circ_0088036 expression was elevated in the synovial lining layer tissues of RA patients and TNF-α-treated RA-FLSs. Circ_0088036 interference largely reversed TNF-α-induced proliferation and inflammation in RA-FLSs. The interaction between circ_0088036 and miR-326 was verified, and miR-326 silencing largely reversed circ_0088036 knockdown-mediated effects in TNF-α-treated RA-FLSs. MiR-326 bound to the 3' untranslated region (3'UTR) of FZD4 in RA-FLSs. FZD4 overexpression largely diminished miR-326 accumulation-mediated influences in TNF-α-treated RA-FLSs. Circ_0088036 could up-regulate FZD4 by sponging miR-326 in RA-FLSs.
Circ_0088036 contributed to TNF-α-induced RA progression partly by targeting miR-326/FZD4 signalling.
环状RNA（circRNAs）在类风湿关节炎（RA）等自身免疫性疾病中的功能及病理意义尚不清楚。在此，我们探讨了circ_0088036在RA进展中的作用及其相关机制。
收集RA患者和非RA对照患者的滑膜衬里层组织用于临床研究，并使用肿瘤坏死因子α（TNF-α）诱导的RA成纤维样滑膜细胞（RA-FLSs）进行实验。通过细胞计数试剂盒8（CCK8）检测和流式细胞术评估细胞增殖。通过流式细胞术分析细胞凋亡。进行酶联免疫吸附测定（ELISA）以分析促炎细胞因子的释放。进行双荧光素酶报告基因检测和RNA免疫沉淀（RIP）检测以验证微小RNA-326（miR-326）与circ_0088036或卷曲蛋白4（FZD4）之间的靶向相互作用。
circ_0088036在RA患者的滑膜衬里层组织和TNF-α处理的RA-FLSs中表达升高。circ_0088036干扰在很大程度上逆转了TNF-α诱导的RA-FLSs增殖和炎症。验证了circ_0088036与miR-326之间的相互作用，并且miR-326沉默在很大程度上逆转了circ_0088036敲低介导的对TNF-α处理的RA-FLSs的影响。在RA-FLSs中，miR-326与FZD4的3'非翻译区（3'UTR）结合。FZD4过表达在很大程度上减弱了miR-326积累介导的对TNF-α处理的RA-FLSs的影响。在RA-FLSs中，circ_0088036可通过海绵吸附miR-326上调FZD4。
circ_0088036部分通过靶向miR-326/FZD4信号通路促进TNF-α诱导的RA进展。
