Altered expression of microRNA‐203 in rheumatoid arthritis synovial fibroblasts and its role in fibroblast activation

J Stanczyk, C Ospelt, E Karouzakis, A Filer… - Arthritis & …, 2011 - Wiley Online Library
J Stanczyk, C Ospelt, E Karouzakis, A Filer, K Raza, C Kolling, R Gay, CD Buckley, PP Tak
Arthritis & Rheumatism, 2011Wiley Online Library
Objective MicroRNA (miRNA) are recognized as important regulators of a variety of
fundamental biologic processes. Previously, we described increased expression of miR‐155
and miR‐146a in rheumatoid arthritis (RA) and showed a repressive effect of miR‐155 on
matrix metalloproteinase (MMP) expression in RA synovial fibroblasts (RASFs). The present
study was undertaken to examine alterations in expression of miR‐203 in RASFs and
analyze its role in fibroblast activation. Methods Differentially expressed miRNA in RASFs …
Objective
MicroRNA (miRNA) are recognized as important regulators of a variety of fundamental biologic processes. Previously, we described increased expression of miR‐155 and miR‐146a in rheumatoid arthritis (RA) and showed a repressive effect of miR‐155 on matrix metalloproteinase (MMP) expression in RA synovial fibroblasts (RASFs). The present study was undertaken to examine alterations in expression of miR‐203 in RASFs and analyze its role in fibroblast activation.
Methods
Differentially expressed miRNA in RASFs versus osteoarthritis synovial fibroblasts (OASFs) were identified by real‐time polymerase chain reaction (PCR)–based screening of 260 individual miRNA. Transfection of miR‐203 precursor was used to analyze the function of miR‐203 in RASFs. Levels of interleukin‐6 (IL‐6) and MMPs were measured by real‐time PCR and enzyme‐linked immunosorbent assay. RASFs were stimulated with IL‐1β, tumor necrosis factor α (TNFα), lipopolysaccharide (LPS), and 5‐azacytidine (5‐azaC). Activity of IκB kinase 2 was inhibited with SC‐514.
Results
Expression of miR‐203 was higher in RASFs than in OASFs or fibroblasts from healthy donors. Levels of miR‐203 did not change upon stimulation with IL‐1β, TNFα, or LPS; however, DNA demethylation with 5‐azaC increased the expression of miR‐203. Enforced expression of miR‐203 led to significantly increased levels of MMP‐1 and IL‐6. Induction of IL‐6 by miR‐203 overexpression was inhibited by blocking of the NF‐κB pathway. Basal expression levels of IL‐6 correlated with basal expression levels of miR‐203.
Conclusion
The current results demonstrate methylation‐dependent regulation of miR‐203 expression in RASFs. Importantly, they also show that elevated levels of miR‐203 lead to increased secretion of MMP‐1 and IL‐6 via the NF‐κB pathway and thereby contribute to the activated phenotype of synovial fibroblasts in RA.
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