circ_0025908 通过调控 miR-650 依赖的 SCUBE2 抑制成纤维样滑膜细胞增殖、迁移、侵袭和炎症，同时促进其凋亡。
Knockdown of circ_0025908 inhibits proliferation, migration, invasion, and inflammation while stimulates apoptosis in fibroblast-like synoviocytes by regulating miR-650-dependent SCUBE2.
机构信息
Department of Rheumatology and Immunology, Xingtai People's Hospital, Xingtai, Hebei Province, China.
Department of Anesthesiology, Weifang People's Hospital, Weifang, China.
出版信息
Autoimmunity. 2022 Nov;55(7):473-484. doi: 10.1080/08916934.2022.2102164. Epub 2022 Jul 29.
BACKGROUND
Circular RNAs (circRNAs) are demonstrated to play vital roles in human diseases, including rheumatoid arthritis (RA). Therefore, this research aimed to explore the effects of hsa_circRNA_0025908 (circ_0025908) on RA.
METHODS
RNA expression of circ_0025908, microRNA-650 (miR-650), and Signal peptide-CUBepidermal growth factor-like containing protein 2 (SCUBE2) were assessed by real-time quantitative polymerase chain reaction; protein expression of SCUBE2, apoptosis- and invasion-related proteins was evaluated by western blot assay. Functional assays were performed using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-2H-tetrazol-3-ium bromide, 5-ethynyl-2'-deoxyuridine, transwell, flow cytometry, and enzyme linked immunosorbent assay assays. Dual-luciferase reporter, RNA immunoprecipitation, and RNA pull-down assays confirmed the interaction relationship among circ_0025908, miR-650, and SCUBE2.
RESULTS
Circ_0025908 was overexpressed in synovial tissues and fibroblast-like synoviocytes (FLS) from RA patients. Inhibition of circ_0025908 repressed proliferation, migration, invasion, inflammation, and cell cycle progression, while induced apoptosis in the FLS isolated from RA patients (FLS-RA), accompanied with increased Bax, cleaved caspase-3 and E-cadherin, but declined Bcl-2, N-cadherin and Vimentin. MiR-650 was a target of circ_0025908, and SCUBE2 was a target for miR-650. Silencing of miR-650 could overturned above effects of circ_0025908 knockdown in FLS-RA, whereas its overexpression could mimic those effects by downregulating SCUBE2. Additionally, SCUBE2 expression could be positively regulated by circ_0025908 and inversely regulated by miR-650. Notably, Pearson's correlation analysis confirmed the linear correlation among circ_0025908, miR-650 and SCUBE2 in these RA tissues.
CONCLUSION
Circ_0025908 inhibition can suppress FLS-RA dysfunctions through targeting miR-650/SCUBE2 axis, suggesting a new potential therapeutic clue for RA patients.
背景
环状 RNA（circRNAs）被证明在包括类风湿关节炎（RA）在内的人类疾病中发挥着重要作用。因此，本研究旨在探讨 hsa_circRNA_0025908（circ_0025908）对 RA 的影响。
方法
通过实时定量聚合酶链反应评估 circ_0025908、微小 RNA-650（miR-650）和信号肽-CUB 表皮生长因子样蛋白 2（SCUBE2）的 RNA 表达；通过 Western blot 检测 SCUBE2、凋亡和侵袭相关蛋白的蛋白表达。使用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基-2H-四唑-3-溴化物（MTT）、5-乙炔基-2'-脱氧尿苷（EdU）、Transwell、流式细胞术和酶联免疫吸附测定（ELISA）试剂盒进行功能测定。通过双荧光素酶报告基因、RNA 免疫沉淀（RIP）和 RNA 下拉实验证实了 circ_0025908、miR-650 和 SCUBE2 之间的相互作用关系。
结果
circ_0025908 在 RA 患者的滑膜组织和成纤维样滑膜细胞（FLS）中高表达。抑制 circ_0025908 可抑制 RA 患者来源的 FLS 的增殖、迁移、侵袭、炎症和细胞周期进程，同时诱导细胞凋亡，伴随着 Bax、cleaved caspase-3 和 E-cadherin 的增加，而 Bcl-2、N-cadherin 和 Vimentin 的减少。miR-650 是 circ_0025908 的靶标，而 SCUBE2 是 miR-650 的靶标。沉默 miR-650 可以逆转 circ_0025908 敲低在 FLS-RA 中的上述作用，而其过表达可以通过下调 SCUBE2 来模拟这些作用。此外，SCUBE2 的表达可以被 circ_0025908 正向调节，被 miR-650 负向调节。值得注意的是，皮尔逊相关分析证实了这些 RA 组织中 circ_0025908、miR-650 和 SCUBE2 之间的线性相关性。
结论
通过靶向 miR-650/SCUBE2 轴抑制 circ_0025908 可抑制 FLS-RA 功能障碍，为 RA 患者提供了新的潜在治疗线索。
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