Abstract

The role and mechanism of lncRNA XIST (XIST) in the development of rheumatoid arthritis (RA) was explored in this study. RT-qPCRs were performed to detect the expression of XIST and miR-126-3p in synovial tissues and cells. Target gene prediction and luciferase gene reporter assay were used to validate downstream target genes of XIST. MTT assay, EdU staining and Annexin V/PI staining were performed to explore the effects of XIST and miR-126-3p on cell proliferation and apoptosis. Western blotting analysis was used to detect the expression of related proteins. We found that the expression levels of XIST in tissues and cells were significantly higher than that in normal tissues and cells. Down-regulation of XIST could inhibit cell proliferation rate and increase apoptosis rate. Luciferase gene reporter assay showed that miR-126-3p was a downstream target gene of XIST. Overexpression of miR-126-3p significantly inhibited RA-FLS cell proliferation and induced RA-FLS cell apoptosis. In addition, down-regulation of XIST could increase the ratio of caspase-3 and Bax/Bcl-2. In addition, overexpression of miR-126-3p could inhibit the NF-κB signalling pathway by reducing the expression levels of p-p65 and p-IκBα in RA-FLS cells. In conclusion, down-regulation of XIST can inhibit the proliferation of synovial fibroblasts by increasing the expression levels of miR-126-3p/NF-κB, thereby inhibiting the occurrence and development of RA.

摘要

本研究探讨了lncRNA XIST（XIST）在类风湿关节炎（RA）发展中的作用和机制。进行 RT-qPCR 以检测 XIST 和 miR-126-3p 在滑膜组织和细胞中的表达。靶基因预测和荧光素酶基因报告分析用于验证 XIST 的下游靶基因。采用MTT法、EdU染色和Annexin V/PI染色探讨XIST和miR-126-3p对细胞增殖和凋亡的影响。Western印迹分析用于检测相关蛋白的表达。我们发现XIST在组织和细胞中的表达水平明显高于在正常组织和细胞中的表达水平。XIST的下调可抑制细胞增殖率并增加细胞凋亡率。萤光素酶基因报告基因检测显示miR-126-3p是XIST的下游靶基因。miR-126-3p的过表达显着抑制RA-FLS细胞增殖并诱导RA-FLS细胞凋亡。此外，XIST 的下调可以增加 caspase-3 和 Bax/Bcl-2 的比率。此外，过表达 miR-126-3p 可以通过降低 RA-FLS 细胞中 p-p65 和 p-IκBα 的表达水平来抑制 NF-κB 信号通路。综上所述，下调 XIST 可通过增加 miR-126-3p/NF-κB 的表达水平来抑制滑膜成纤维细胞的增殖，从而抑制 RA 的发生和发展。XIST 的下调可增加 caspase-3 和 Bax/Bcl-2 的比率。此外，过表达 miR-126-3p 可以通过降低 RA-FLS 细胞中 p-p65 和 p-IκBα 的表达水平来抑制 NF-κB 信号通路。综上所述，下调 XIST 可通过增加 miR-126-3p/NF-κB 的表达水平来抑制滑膜成纤维细胞的增殖，从而抑制 RA 的发生和发展。XIST 的下调可增加 caspase-3 和 Bax/Bcl-2 的比率。此外，过表达 miR-126-3p 可以通过降低 RA-FLS 细胞中 p-p65 和 p-IκBα 的表达水平来抑制 NF-κB 信号通路。综上所述，下调 XIST 可通过增加 miR-126-3p/NF-κB 的表达水平来抑制滑膜成纤维细胞的增殖，从而抑制 RA 的发生和发展。