Department of Rheumatology, Xi'an No.5 Hospital, Xi'an, China.
Medical Department, Hospital of Northwest Polytechnic University, Xi'an, China.
Autoimmunity. 2022 Dec;55(8):567-576. doi: 10.1080/08916934.2022.2126460. Epub 2022 Sep 26.
Rheumatoid arthritis (RA) is a chronic progressive autoimmune disease of unknown etiology. Human fibroblast-like synoviocytes (HFLSs) are the main effector cells for synovial hyperplasia and invasion in RA. Long non-coding RNAs (lncRNAs) play key roles in several autoimmune diseases, including RA. We investigated the effects of lncRNA HOX transcript antisense intergenic RNA (HOTAIR) on the pathological behavior of HFLSs in RA. The microRNAs (miRNAs) with potential binding sites for lncRNA HOTAIR were predicted using Starbase v2.0. TargetScan (http://www.targetscan.org) was used to analyze the potential target genes of miR-106b-5p. The interactions were further verified using a dual-luciferase reporter assay. RNA and protein expression was determined using quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blotting. The proliferation, cell invasion and migration, and cell apoptosis of HFLSs in RA was detected by the 3-(4,5-dimethylthiazol)-2,5-diphenyl-tetrazolium bromide (MTT) assay, transwell assay, and flow cytometry (FCM). The dual luciferase reporter assay confirmed the interactions between lncRNA HOTAIR and miR-106b-5p and between miR-106b-5p and SMAD family member 7 (SMAD7). The qRT-PCR results indicated that the expression of lncRNA HOTAIR was markedly decreased and that of miR-106b-5p was markedly increased in HFLSs of RA. Cell proliferation, invasion, and migration of HFLSs were inhibited by lncRNA HOTAIR upregulation, and the expression of miR-106b-5p was negatively regulated by lncRNA HOTAIR in HFLSs. Apoptosis of HFLS cells was improved by the overexpression of lncRNA HOTAIR. All the effects of lncRNA HOTAIR upregulation on HFLSs were reversed after the overexpression of miR-106b-5p. Smad7 was identified as a target gene of miR-106b-5p, and the effects of downregulation of miR-106b-5p on HFLSs could be abolished by silencing Smad7. We found that lncRNA HOTAIR was significantly downregulated in the HFLSs of patients with RA. Moreover, lncRNA HOTAIR influenced cell growth, migration, invasion, and apoptosis in HFLSs through the miR-106b-5p/Smad7 axis.
类风湿关节炎（RA）是一种病因不明的慢性进行性自身免疫性疾病。人类成纤维样滑膜细胞（HFLS）是 RA 滑膜过度增生和侵袭的主要效应细胞。长链非编码 RNA（lncRNA）在包括 RA 在内的几种自身免疫性疾病中发挥着关键作用。我们研究了 lncRNA HOX 转录反义基因间 RNA（HOTAIR）对 RA 中 HFLS 病理行为的影响。使用 Starbase v2.0 预测与 lncRNA HOTAIR 具有潜在结合位点的 microRNAs（miRNAs）。使用 TargetScan（http://www.targetscan.org）分析 miR-106b-5p 的潜在靶基因。通过双荧光素酶报告基因实验进一步验证相互作用。使用实时定量逆转录聚合酶链反应（qRT-PCR）和蛋白质印迹法检测 RNA 和蛋白质表达。通过 3-（4,5-二甲基噻唑-2,5-二苯基四唑溴盐）（MTT）测定法、Transwell 测定法和流式细胞术（FCM）检测 RA 中 HFLS 的增殖、细胞侵袭和迁移以及细胞凋亡。双荧光素酶报告基因实验证实了 lncRNA HOTAIR 与 miR-106b-5p 以及 miR-106b-5p 与 SMAD 家族成员 7（SMAD7）之间的相互作用。qRT-PCR 结果表明，RA 中 HFLS 的 lncRNA HOTAIR 表达明显降低，miR-106b-5p 表达明显升高。上调 lncRNA HOTAIR 可抑制 HFLS 的增殖、侵袭和迁移，下调 lncRNA HOTAIR 可负调控 HFLS 中 miR-106b-5p 的表达。上调 lncRNA HOTAIR 可改善 HFLS 细胞的凋亡。过表达 miR-106b-5p 可逆转上调 lncRNA HOTAIR 对 HFLS 的所有作用。SMAD7 被鉴定为 miR-106b-5p 的靶基因，沉默 SMAD7 可消除 miR-106b-5p 对 HFLS 的影响。我们发现 RA 患者的 HFLS 中 lncRNA HOTAIR 明显下调。此外，lncRNA HOTAIR 通过 miR-106b-5p/Smad7 轴影响 HFLS 的细胞生长、迁移、侵袭和凋亡。
