Posttranscriptional regulation of collagenase‐1 gene expression in synoviocytes by adenosine receptor stimulation

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Abstract
Objective. To characterize the transcriptional and posttranscriptional regulation of collagenase‐1 by adenosine receptor stimulation in interleukin‐1 (IL‐1)‐stimulated fibroblast‐like synoviocytes (FLS). Methods. FLS were stimulated with IL‐1 and either the nonselective adenosine agonist 5′‐N‐ethylcarboxamidoadenosine (NECA) or the adenylate cyclase activator forskolin. Electrophoretic mobility shift assays were performed to determine AP‐1 and cAMP‐responsive element binding protein (CREB) activation. Transcriptional activation was determined by transfecting HS68 dermal fibroblasts with a collagenasechloramphenicol acetyltransferase construct. Finally, collagenase messenger RNA (mRNA) half‐life was determined by activating cells in the presence of IL‐1, IL‐1 + NECA, or IL‐1 + forskolin and culturing cells in the presence of actinomycin D. Results. NECA and forskolin had no effect on AP‐1 activation, c‐jun or c‐fos gene expression, or CREB phosphorylation. IL‐1 markedly increased collagenase promoter activity, and neither NECA nor forskolin blocked this action. Studies of mRNA half‐life showed that both NECA and forskolin decreased the half‐life of collagenase mRNA in IL‐1‐stimulated FLS and HS68 cells. Conclusion. The findings of this study demonstrate that NECA and forskolin decrease collagenase gene expression in FLS and dermal fibroblasts due to enhanced mRNA degradation.

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