Long non-coding RNA PVT1 can regulate the proliferation and inflammatory responses of rheumatoid arthritis fibroblast-like synoviocytes by targeting microRNA-145-5p

• Published in: Human cell : official journal of Human Cell Research Society 2020-10, Vol.33 (4), p.1081-1090
• Main Authors: Tang, Jiajun, Yi, Shiyu, Liu, Yi
• Format: Article
• Language: English
• Subjects: Adult; Aged; Apoptosis; Apoptosis - genetics; Arthritis, Rheumatoid - drug therapy; Arthritis, Rheumatoid - genetics; Arthritis, Rheumatoid - pathology; Binding sites; Biomedical and Life Sciences; Cell Biology; Cell proliferation; Cell Proliferation - genetics; Cells, Cultured; Cytokines - metabolism; Enzyme-linked immunosorbent assay; Female; Flow cytometry; Gene Expression; Gynecology; Humans; Inflammation; Inflammation - genetics; Inflammation Mediators - metabolism; Life Sciences; Male; MicroRNAs; MicroRNAs - metabolism; Middle Aged; miRNA; Molecular Targeted Therapy; mRNA; NF-κB protein; Non-coding RNA; Oncology; Polymerase chain reaction; Reproductive Medicine; Research Article; Rheumatoid arthritis; RNA, Long Noncoding - genetics; RNA, Long Noncoding - metabolism; RNA, Long Noncoding - physiology; Stem Cells; Surgery; Synoviocytes; Synoviocytes - metabolism; Synoviocytes - pathology; Tumor Necrosis Factor-alpha; Tumor necrosis factor-TNF; Tumor necrosis factor-α; Tumors
• ISSN: 1749-0774; 0914-7470; 1749-0774
• DOI: 10.1007/s13577-020-00419-6

Long non-coding RNAs (lncRNAs) function in rheumatoid arthritis (RA). The present work was designed to explore the roles of lncRNA PVT1 in RA and the related mechanism. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to determine mRNA level. The binding sites between PVT1 and miR-145-5p were verified by a dual-luciferase reporter assay. Furthermore, RA-FLSs were treated with TNF-α to establish the RA model. 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and 5-ethynyl-2′-deoxyuridine (EdU) assays were performed to detect cell proliferation. Flow cytometry and TUNEL assays were performed to detect cell apoptosis. Enzyme-linked immunosorbent assay (ELISA) was used to determine levels of inflammatory cytokines. PVT1 was significantly increased and miR-145-5p was decreased in synovial tissues of RA patients. miR-145-5p is a target miRNA of PVT1, and the levels of PVT1 and miR-145-5p in synovial tissues of RA patients were negatively correlated. In RA-FLSs, tumour necrosis factor-α (TNF-α) led to increased PVT1 levels and decreased miR-145-5p levels. Knockdown of PVT1 inhibited TNF-α-induced RA-FLS over-proliferation and reversed TNF-α-induced RA-FLS apoptosis reduction. Moreover, knockdown of PVT1 inhibited TNF-α-induced production of interleukin (IL)‐1β and IL‐6 and the activation of NF-κB through miR-145-5p. PVT1 can regulate apoptosis and inflammatory responses in RA-FLSs by targeting miR-145-5p.