Hypoxia-induced miR-191-C/EBPβ signaling regulates cell proliferation and apoptosis of fibroblast-like synoviocytes from patients with rheumatoid arthritis
Hypoxia-induced miR-191-C/EBPβ signaling regulates cell proliferation and apoptosis of fibroblast-like synoviocytes from patients with rheumatoid arthritis
复制标题
缺氧诱导的 miR-191-C/EBP beta 信号传导调节类风湿性关节炎患者成纤维样滑膜细胞的细胞增殖和凋亡
DOI：
10.1186/s13075-019-1861-7
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发表时间：
2019-03-20
影响因子：
4.9
通讯作者：
Fan, Lieying
中科院分区：
文献类型：
--
作者：
Yu, Shanshan;Lu, Ying;Fan, Lieying
背景：低氧在类风湿关节炎成纤维细胞样滑膜细胞(RA-FLS)的增殖中起重要作用，导致类风湿关节炎的病理改变。本研究旨在探讨低氧诱导的RA-FLS中的microRNAs(LowxamiR)及其在RA-FL功能中的作用。方法：在常氧(21%O-2)和低氧(3%O-2)条件下培养RA-FLS，并进行microRNA(MiRNA)阵列分析。实时定量聚合酶链式反应(RT-PCR)证实，低氧可使RA-FLS和骨性关节炎(OA)患者的FLS中miR-191表达上调。通过对miR-191模拟物和抑制物的转染，研究miR-191在RA-FLS中的作用。利用生物信息学对miR-191的功能靶点进行预测，并通过报告基因分析对其进行验证。结果：鉴定出包括miR-191在内的一组miRNAs受缺氧诱导。与缺氧性OA-FLS相比，miR-191的上调在缺氧性RA-FLS中具有特异性。我们观察到，在RA-FLS中，miR-191通过促进细胞周期的G(1)/S转变而促进细胞增殖，并抑制细胞饥饿诱导的细胞凋亡。生物信息学分析和实验分析证实CCAAT/增强子结合蛋白β(C/EBPβ)是RA-FLS中miR-191的靶基因。增强C/EBPβ的表达挽救了miR-191诱导的细胞表型。此外，在RA-FLS中，C/EBPβ水平与低氧刺激呈负相关，C/EBPβ的过度表达可以部分挽救低氧诱导的细胞增殖。结论：我们证实了miR-191-C/EBPβ信号通路介导了低氧诱导的RA细胞增殖。
Background: Hypoxia plays an important role in the proliferation of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS), leading to pathology of RA. This study was conducted to evaluate hypoxia-induced microRNAs (hypoxamiR) in RA-FLS and its role in the function of RA-FLS.Methods: RA-FLS were cultured under normoxia (21% O-2) or hypoxia (3% O-2) condition, followed by a microRNA (miRNA) array analysis. The upregulation of miR-191 by hypoxia was confirmed in RA-FLS and FLS from osteoarthritis (OA) patients by quantitative real-time polymerase chain reaction (RT-PCR). Transfection of miR-191 mimic and inhibitor was used to investigate the function of miR-191 in RA-FLS. The functional targets of miR-191 were predicted by bioinfomatics and then validated by reporter gene assay.Results: A subset of miRNAs was identified to be induced by hypoxia including miR-191. The upregulation of miR-191 was found to be specific in hypoxic RA-FLS, compared to hypoxic OA-FLS. We observed that miR-191 in RA-FLS increased cellular proliferation via promoting G(1)/S transition of the cell cycle and suppressed cell apoptosis induced by cell starvation. Bioinformatical analysis and experimental assays identified CCAAT/enhancer binding protein beta (C/EBP beta) as a target gene of miR-191 in RA-FLS. Enforced expression of C/EBP beta rescued the cellular phenotypes induced by miR-191. In addition, an inverse correlation between the C/EBP beta level and hypoxia stimulation was found in RA-FLS, and overexpression of C/EBP beta could partly rescue the hypoxia-induced cell proliferation.Conclusion: We demonstrated the miR-191-C/EBP beta signaling pathway mediating the hypoxia-induced cell proliferation in RA.