MicroRNA-495 attenuates proliferation and inflammatory response in rheumatoid arthritis fibroblast-like synoviocytes through attenuating β-catenin pathway
Fibroblast like synoviocytes (FLSs) exert important effects in the occurrence and progression of rheumatoid arthritis (RA). MicroRNA-495 (miR-495) can regulate growth behavior of many cell types. Nevertheless, the role of miR-495 is still unclear in RA-FLS. We aim to explore the role and molecular mechanism of miR-495 in RA. The FLSs and synovial tissue from normal and RA cases were used in this study. RT-PCR analysis was used to test the expression of miR-495. Western blot assay was performed to determine the levels of matrix metalloproteinase-9 (MMP-9), matrix metalloproteinase-2 (MMP-2) and β-catenin.
Cell counting kit-8 (CCK-8) tests were performed to determine the proliferation of RA-FLS in the different treatment groups. The results showed that miR-495 down-regulated in the synovial tissue and RA-RA-FLSs. Overexpression of miR-495 can inhibit RA-FLS proliferation and inflammatory factor interleukin (IL) -6, IL-11 and tumor necrosis factor alpha (TNF-α), and the lower the protein expression of MMP-9 and MMP-2. In addition, miR-495 could negatively regulates the expression of β-catenin in RA-FLSs.
We also confirmed that the role of inhibition of miR-495 in RA-FLS is through the regulation of β-catenin expression. Taken together, miR-495 is downregulated in RA-FLS and RA synovial tissue, and miR-495 inhibits proliferation and the inflammatory response in RA-FLS, partly through regulating the expression of β-catenin. The miR-495 / β-catenin signaling pathway may serve as a new therapeutic target for RA. mir-483-3p was upregulated in RA, which is explicitly promoted cell proliferation, induction of the transition phase G0 / G1-to-S, and suppressed apoptosis in RA FLSs, whereas silencing of miR-483-3p produce the opposite result.
In addition, insulin growth factor 1 (IGF-1) is detected as a direct target of miR-483-3p. IGF-1 silencing partially restored cell proliferation, the transition phase G0 / G1-to-S, and suppression of apoptosis in RA FLSs through inhibition of miR-483-3p. Our results show that miR-483-3p RA FLSs promote proliferation by targeting the IGF-1, indicating a potential strategy for diagnostic and treatment strategies for RA.
MicroRNA-495 attenuates proliferation and inflammatory response in rheumatoid arthritis fibroblast-like synoviocytes through attenuating β-catenin pathway
MicroRNA-15a / 16 / SOX5 axis promotes migration, invasion and the inflammatory response in rheumatoid arthritis fibroblast like synoviocytes
Fibroblast like synoviocytes (FLSs) is a key effector cells in the pathogenesis of rheumatoid arthritis (RA) and featuring aggressive tumor phenotype is unique with exceptional hyperplasia, increased cell migration and invasion. How FLSs experiencing this change in RA remains unknown. We previously reported a novel function of transcription factors in RA SOX5-FLSs that promote cell migration and invasion.
In this study, we found that miR-15a / 16 directly target and suppress the expression of the 3’UTR SOX5 SOX5. In addition, miR-15a / 16 was significantly down-regulated in RA-FLSs, which is negatively correlated with the expression SOX5. Transfection with miR-15a / 16 mimic the RA-FLSs cell migration inhibiting, invasion, IL-1β and TNF expression. SOX5 expression in RA-FLSs decreased miR-15a / 16 expression and rescue miR-15a / 16-mediated inhibitory effect.
Description: CD300C Human Recombinant produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing 163 amino acids (29-183a.a.) and having a molecular mass of 18kDa (Molecular size on SDS-PAGE will appear at approximately 28-40 kDa). CD300C is expressed with an 8 amino acid His tag at C-Terminus and purified by proprietary chromatographic techniques.
Description: Noggin Human Recombinant produced in HEK cells is a glycosylated homodimer, having a total molecular weight of 65kDa.;The Noggin is purified by proprietary chromatographic techniques.
Description: Noggin produced in Sf9 Insect cells is a single, glycosylated polypeptide chain containing 211 amino acids (28-232a.a.) and having a molecular mass of 23.8kDa. (Molecular size on SDS-PAGE will appear at approximately 28-40kDa).; Noggin is expressed with a 6 amino acid His tag at C-Terminus and purified by proprietary chromatographic techniques.
Description: SLAMF1 Human Recombinant produced in Sf9 Baculovirus cells is a single, glycosylated polypeptide chain containing 226 amino acids (21-237a.a.) and having a molecular mass of 25.3kDa (Molecular size on SDS-PAGE will appear at approximately 28-40kDa).;SLAMF1 is expressed with a 6 amino acid His tag at C-Terminus and purified by proprietary chromatographic techniques.
Description: Periostin Human Recombinant produced in HEK cells is a single, glycosylated, polypeptide chain (Asn22-Gln836) containing a total of 821 amino acids, having a calculated molecular mass of 91.8kDa and fused to a 6 aa His tag at C-Terminus.
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Furthermore, RA patients with low baseline serum miR-15a / 16 levels now the poor response of the 3-month disease-modifying antirheumatic drugs (DMARDs) therapy. Collectively, these studies revealed that miR-15a / 16 / SOX5 axis serves as the main driver of RA-FLSs invasion, migration and inflammatory responses in each negative feedback and correlated with treatment response to DMARDs in RA.
Shelby
