Effects of miR-150-5p on the growth and SOCS1 expression of rheumatoid arthritis synovial fibroblasts

Objective miR-150-5p has been implicated in the regulation and onset of immune diseases. We investigated the effects of miR-150-5p on the functions of RA synovial fibroblasts (RASFs). Method The binding site between suppressor of cytokine signaling 1 (SOCS1) and miR-150-5p was analyzed using Europea...

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Veröffentlicht in:Clinical rheumatology 2020-03, Vol.39 (3), p.909-917
Hauptverfasser: Qiu, Mingliang, Mo, Lisha, Li, Juxiang, Liang, Hua, Zhu, Weina, Zheng, Xiangjuan, Duan, Xinwang, Xu, Weidong
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Sprache:eng
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Zusammenfassung:Objective miR-150-5p has been implicated in the regulation and onset of immune diseases. We investigated the effects of miR-150-5p on the functions of RA synovial fibroblasts (RASFs). Method The binding site between suppressor of cytokine signaling 1 (SOCS1) and miR-150-5p was analyzed using European Bioinformatics Institute database, and the 3′ UTR of SOCS1 mRNA, including the binding site, was amplified and ligated to the 3′-end of LUC2 gene in the pmirGL0 dual-luciferase vector. The pmirGL0 vector and corresponding mimics were subsequently co-transfected into 293T cells to compare the relative fluorescence intensity of LUC2 between the miR-150-5p mimics and the negative control (NC) mimics groups. Further, the RASF cell line MH7A was transfected with miR-150-5p or NC mimics and subjected to flow cytometric analysis, cell counting kit-8 assay, western blot analysis, qPCR, and enzyme-linked immunosorbent (ELISA) assay 48 h after transfection. Results miR-150-5p mimics resulted in a lower cell apoptotic rate and proportion of cells in the S phase. Using a dual-luciferase reporter gene assay, we then found that SOCS1 is a potential target of miR-150-5p. Compared with NC mimics, miR-150-5p mimics significantly decreased the protein and mRNA expression levels of SOCS1. ELISA assay showed that miR-150-5p mimics increased interleukin-6 level in the cell culture medium but did not influence tumor necrosis factor-alpha levels. Conclusions Overall, the growth-promoting effect of miR-150-5p on MH7A cells may be attributed to the miR-150-5p-induced degradation of SOCS1 mRNA, suggesting a potential therapeutic target for RA. Key Points • SOCS1 is a potential target of miR-150-5p . • miR-150-5p promoted the growth of RASF cell line MH7A . • miR-150-5p increased the secretion of IL-6 but did not significantly affect TNF-α levels in MH7A cells .
ISSN:0770-3198
1434-9949
DOI:10.1007/s10067-019-04894-7