Wensi Su, Zhifang Ye, Guangji Wang, Hui Huang, Yehan Fang
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引用次数: 0
摘要
环状 RNA(circRNA)在类风湿性关节炎(RA)的发展过程中发挥着功能性作用。成纤维细胞样滑膜细胞(RASFs)是类风湿性关节炎发展过程中的主要效应因子。本研究探讨了circ_0008410在RASFs中的功能和机制。采用qRT-PCR检测circ_0008410、microRNA-149-5p(miR-149-5p)和homeodomain-interacting protein kinase 2(HIPK2)的表达。细胞计数试剂盒-8、EdU 试验、流式细胞术和透孔试验用于评估细胞增殖、凋亡、迁移和侵袭。Western blot 检测了相关标志物和 HIPK2 的蛋白水平。通过相应的酶联免疫吸附试剂盒和 Western blot 检测了 IL-1β、TNF-α 和 IL-6 的水平。miR-149-5p与circ_0008410或HIPK2的结合通过双荧光素酶报告实验或RNA免疫沉淀(RIP)实验进行检测。我们的数据显示,在RA滑膜组织和RASFs中,circ_0008410和HIPK2升高,而miR-149-5p下调。Circ_0008410促进了RASF的增殖、迁移、侵袭和炎症反应,同时抑制了细胞凋亡。miR-149-5p是circ_0008410的靶标,其过表达可逆转circ_0008410对RASF功能障碍的促进作用。此外,miR-149-5p 还能靶向 HIPK2,抑制 RASF 的增殖、迁移、侵袭和炎症。总之,circ_0008410通过miR-149-5p/HIPK2促进了RASF的功能障碍,这可能为RA的治疗提供了一个潜在的靶点。
Circ_0008410 contributes to fibroblast-like synoviocytes dysfunction by regulating miR-149-5p/HIPK2 axis
Circular RNAs (circRNAs) play functional roles in rheumatoid arthritis (RA) progression. Fibroblast-like synoviocytes (RASFs) are the main effectors in RA development. In this study, we explored the function and mechanism of circ_0008410 in RASFs. qRT-PCR was used to detect the expression of circ_0008410, microRNA-149-5p (miR-149-5p), and homeodomain-interacting protein kinase 2 (HIPK2). Cell counting kit-8, EdU assay, flow cytometry, and transwell assay were performed to evaluate cell proliferation, apoptosis, migration, and invasion. Western blot measured the protein levels of related markers and HIPK2. The levels of IL-1β, TNF-α, and IL-6 were tested by corresponding ELISA kits and Western blot. The combination between miR-149-5p and circ_0008410 or HIPK2 was detected by dual-luciferase reporter assay or RNA immunoprecipitation (RIP) assay. Our data showed that circ_0008410 and HIPK2 were elevated, while miR-149-5p was downregulated in RA synovial tissues and RASFs. Circ_0008410 promoted RASF proliferation, migration, invasion, and inflammation while inhibiting apoptosis. MiR-149-5p was a target of circ_0008410, and its overexpression could reverse the promoting effects of circ_0008410 on RASF dysfunction. Moreover, miR-149-5p could target HIPK2 to suppress RASF proliferation, migration, invasion, and inflammation. Collectively, circ_0008410 promoted RASF dysfunction via miR-149-5p/HIPK2, which might provide a potential target for RA therapy.
期刊介绍:
Microbiology and Immunology is published in association with Japanese Society for Bacteriology, Japanese Society for Virology, and Japanese Society for Host Defense Research. It is peer-reviewed publication that provides insight into the study of microbes and the host immune, biological and physiological responses.
Fields covered by Microbiology and Immunology include:Bacteriology|Virology|Immunology|pathogenic infections in human, animals and plants|pathogenicity and virulence factors such as microbial toxins and cell-surface components|factors involved in host defense, inflammation, development of vaccines|antimicrobial agents and drug resistance of microbes|genomics and proteomics.