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: Human synoviocytes (HS), the predominant cell type of healthy synovial tissue, are fibroblast-like cells. The Synoviocytes form a distinct structure called the synovial lining layer. Electron microscopy revealed extensive cell-to-cell contacts within the lining layer. The Synoviocytes produce synovial fluid components and are responsible for absorption from the joint cavity, and for blood/synovial fluid exchanges. The synoviocytes proliferate, show anchorage-independent growth, and also secrete a variety of effector molecules that promote inflammation and joint destruction and, themselves are part of a complex network of autocrine and paracrine acting factors. The synoviocytes express cadherin-11 providing new insight into synovial tissue organization and morphogenesis and of interest as a therapeutic target.HS from Gentaur Research Laboratories are isolated from human synovium. HS are cryopreserved at passage one cultures and delivered frozen. Each vial contains 5×10^5 cells in 1 ml volume. HS are characterized by their fibroblast-like morphology, growth pattern and immunocytochemistry of CD 90 and fibronectin. HS are negative for HIV-1, HBV, HCV, mycoplasma, bacteria, yeast and fungi. HS are guaranteed for 15 population doublings at the conditions provided by Gentaur Research Laboratories.
Description: Human Fibroblast-Like Synoviocytes (HFLS) are isolated from normal synovial tissues. They are cryopreserved at second passage and can be cultured and propagated at least 5 population doublings.
Description: Human Fibroblast-Like Synoviocytes (HFLS) are isolated from synovial tissues obtained from patients with Rheumatoid Arthritis (RA) / Osteoarthritis (OA). They are cryopreserved at second passage and can be cultured and propagated at least 5 population doubling.
Synoviocyte Growth Medium, Ready-to-Use, 500 ml, 10% Human Serum Supplement
Description: Human Fibroblast-Like Synoviocytes (HFLS) are isolated from synovial tissues obtained from patients with Rheumatoid Arthritis (RA) / Osteoarthritis (OA). They are cryopreserved at second passage and can be cultured and propagated at least 5 population doubling.
COOLCELL SV2 STEMCELL CRYOPRESERVATION SYSTEM INCLUDES THE XT STARTER BENCHTOP COOLER, COOLRACK SV2 AND COOLCELL SV2 CELL FREEZING CONTAINER FOR 12 INJECTABLE VIALS
COOLCELL SV10 STEMCELL CRYOPRESERVATION SYSTEM INCLUDES THE XT STARTER BENCHTOP COOLER, COOLRACK SV10 AND COOLCELL SV10 CELL FREEZING CONTAINER FOR 6 INJECTABLE VIALS
Description: Quantitative sandwich ELISA for measuring Human E3 ubiquitin-protein ligase synoviolin (SYVN1) in samples from cell culture supernatants, serum, whole blood, plasma and other biological fluids.
Recombinant Human E3 ubiquitin-protein ligase synoviolin (SYVN1)
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
