Co-activation of synovial fibroblasts by laminin-111 and transforming growth factor-beta induces expression of matrix metalloproteinases 3 and 10 independently of nuclear factor-kappaB

Ann Rheum Dis. 2008 Apr;67(4):559-62. doi: 10.1136/ard.2007.073809. Epub 2007 Aug 24.

Abstract

We showed previously that the attachment of synovial fibroblasts to laminin (LM)-111 in the presence of transforming growth factor-beta induces significant expression of the matrix metalloproteinase (MMP)-3. Here we go on to investigate the regulation of additional MMPs and their specific tissue inhibitors of matrix proteases (TIMPs). Changes in steady-state mRNA levels encoding TIMPs and MMPs were investigated by quantitative reverse transcription-polymerase chain reaction. Production of MMPs was monitored by a multiplexed immunoarray. Signal transduction pathways were studied by immunoblotting. Attachment of synovial fibroblasts to LM-111 in the presence of transforming growth factor-beta induced significant increases in MMP-3 mRNA (12.35-fold, p < 0.001) and protein (mean 62 ng/ml, sixfold, p < 0.008) and in expression of MMP-10 mRNA (11.68-fold, p < 0.05) and protein (54 ng/ml, 20-fold, p > or = 0.02). All other TIMPs and MMPs investigated failed to show this LM-111-facilitated transforming growth factor-beta response. No phosphorylation of nuclear factor-kappaB was observed. We conclude that co-stimulation of synovial fibroblasts by LM-111 together with transforming growth factor-beta suffices to induce significant expression of MMP-3 and MMP-10 by synovial fibroblasts and that this induction is independent of nuclear factor-kappaB phosphorylation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arthritis, Rheumatoid / enzymology*
  • Arthritis, Rheumatoid / metabolism
  • Arthritis, Rheumatoid / pathology
  • Cells, Cultured
  • Early Growth Response Protein 1 / metabolism
  • Electrophoresis, Polyacrylamide Gel / methods
  • Fibroblasts / drug effects*
  • Fibroblasts / enzymology
  • Fibroblasts / metabolism
  • Gene Expression Regulation / drug effects
  • Humans
  • Laminin / pharmacology
  • Matrix Metalloproteinase 10 / biosynthesis*
  • Matrix Metalloproteinase 10 / genetics
  • Matrix Metalloproteinase 3 / biosynthesis*
  • Matrix Metalloproteinase 3 / genetics
  • NF-kappa B / metabolism
  • Osteoarthritis / enzymology
  • Osteoarthritis / metabolism
  • Osteoarthritis / pathology
  • Phosphorylation
  • Proto-Oncogene Proteins c-fos / metabolism
  • RNA, Messenger / genetics
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Synovial Membrane / drug effects
  • Synovial Membrane / enzymology
  • Synovial Membrane / metabolism
  • Tissue Inhibitor of Metalloproteinases / biosynthesis
  • Tissue Inhibitor of Metalloproteinases / genetics
  • Transforming Growth Factor beta / pharmacology

Substances

  • EGR1 protein, human
  • Early Growth Response Protein 1
  • Laminin
  • NF-kappa B
  • Proto-Oncogene Proteins c-fos
  • RNA, Messenger
  • Tissue Inhibitor of Metalloproteinases
  • Transforming Growth Factor beta
  • laminin 1
  • Matrix Metalloproteinase 3
  • Matrix Metalloproteinase 10