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...
Gespeichert in:
Veröffentlicht in: | Clinical rheumatology 2020-03, Vol.39 (3), p.909-917 |
---|---|
Hauptverfasser: | , , , , , , , |
Format: | Artikel |
Sprache: | eng |
Schlagworte: | |
Online-Zugang: | Volltext |
Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
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 |