Effects of miR-150-5p on the growth and SOCS1 expression of rheumatoid arthritis synovial fibroblasts.
Clin Rheumatol
; 39(3): 909-917, 2020 Mar.
Article
in En
| MEDLINE
| ID: mdl-31879859
ABSTRACT
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.Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Arthritis, Rheumatoid
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Synovial Membrane
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MicroRNAs
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Fibroblasts
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Suppressor of Cytokine Signaling 1 Protein
Limits:
Humans
Language:
En
Journal:
Clin Rheumatol
Year:
2020
Document type:
Article