MicroRNA-449 靶向组蛋白去乙酰化酶 1 调节类风湿关节炎滑膜成纤维细胞的增殖、侵袭和凋亡。
MicroRNA-449 targets histone deacetylase 1 to regulate the proliferation, invasion, and apoptosis of synovial fibroblasts in rheumatoid arthritis.
机构信息
Department of Rheumatology and Immunology, People's Hospital of Ningxia Hui Autonomous Region, Yinchuan, China; The Third Affiliated Hospital, Ningxia Medical University, Yinchuan, China; The First Affiliated Hospital of Northwest Minzu University, Yinchuan, China; Ningxia Key Laboratory of Precision Medicine for Autoimmune Diseases, Yinchuan, China.
The Third Affiliated Hospital, Ningxia Medical University, Yinchuan, China; The First Affiliated Hospital of Northwest Minzu University, Yinchuan, China; Department of Neurology, People's Hospital of Ningxia Hui Autonomous Region, Yinchuan, China.
出版信息
Ann Palliat Med. 2021 Jul;10(7):7960-7969. doi: 10.21037/apm-21-1383.
BACKGROUND
Rheumatoid arthritis (RA) is a chronic joint disease. The study aimed to explore the effects of microRNA (miR)-449 and histone deacetylase 1 (HDAC1) on the proliferation, invasion, and apoptosis of synovial fibroblasts in rheumatoid arthritis.
METHODS
Synovial tissue was collected from 20 patients with RA and 20 patients with osteoarthritis (OA) who underwent joint replacement surgery. RA synovial fibroblasts (RASFs) and OA synovial fibroblasts (OASFs) were isolated and cultured. Real-time quantitative PCR was used to detect the expression levels of miR-449 and HDAC1 in synovial tissues and cells. Western blot was performed to detect the cellular expression levels of HDAC1 protein, and apoptosis and invasion-related proteins. The proliferation, invasion, and apoptosis of RASFs were detected by MTT assay, Transwell assay, and flow cytometry. The dual-luciferase reporter gene was used to test the targeting relationship between inflammatory miR-449 and HDAC1.
RESULTS
Compared with normal synovial tissue and OASFs, the levels of HDAC1 messenger RNA in RA synovial tissue and RASF cells were significantly increased (P<0.01), while the expression levels of miR-449 were significantly decreased (P<0.01). The dual-luciferase reporter gene experiment confirmed that miR-449 could specifically bind to the 3' untranslated region of HDAC1 to inhibit its luciferase activity (P<0.05). HDAC1 inhibition or miR-449 overexpression significantly inhibited the proliferation and invasion of RASFs (P<0.001), while inducing their apoptosis (P<0.001). HDAC1 overexpression reversed the biological effects of miR-449 on RASFs (P<0.001).
CONCLUSIONS
miR-449 inhibits the proliferation and invasion of RASFs and induces their apoptosis by targeting HDAC1, thereby exerting a protective effect against RA.
背景
类风湿关节炎（RA）是一种慢性关节疾病。本研究旨在探讨微小 RNA（miR）-449 和组蛋白去乙酰化酶 1（HDAC1）对类风湿关节炎滑膜成纤维细胞增殖、侵袭和凋亡的影响。
方法
收集 20 例 RA 和 20 例接受关节置换术的骨关节炎（OA）患者的滑膜组织。分离并培养 RA 滑膜成纤维细胞（RASFs）和 OA 滑膜成纤维细胞（OASFs）。实时定量 PCR 检测滑膜组织和细胞中 miR-449 和 HDAC1 的表达水平。Western blot 检测 HDAC1 蛋白及凋亡和侵袭相关蛋白的细胞表达水平。MTT 法、Transwell 法和流式细胞术检测 RASFs 的增殖、侵袭和凋亡。双荧光素酶报告基因检测炎症 miR-449 与 HDAC1 的靶向关系。
结果
与正常滑膜组织和 OASFs 相比，RA 滑膜组织和 RASF 细胞中的 HDAC1 信使 RNA 水平明显升高（P<0.01），而 miR-449 的表达水平明显降低（P<0.01）。双荧光素酶报告基因实验证实 miR-449 可特异性结合 HDAC1 的 3'UTR 抑制其荧光素酶活性（P<0.05）。HDAC1 抑制或 miR-449 过表达显著抑制 RASFs 的增殖和侵袭（P<0.001），同时诱导其凋亡（P<0.001）。HDAC1 过表达逆转了 miR-449 对 RASFs 的生物学效应（P<0.001）。
结论
miR-449 通过靶向 HDAC1 抑制 RASFs 的增殖和侵袭并诱导其凋亡，从而对 RA 发挥保护作用。
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