Molecular mechanism of the induction of metalloproteinases 1 and 3 in human fibroblasts by basic calcium phosphate crystals. Role of calcium-dependent protein kinase C alpha

J Biol Chem. 2002 Apr 26;277(17):15190-8. doi: 10.1074/jbc.M200278200. Epub 2002 Feb 8.

Abstract

Synovial fluid basic calcium phosphate (BCP) crystals are common in osteoarthritis and are often associated with destructive arthropathies involving cartilage degeneration. These crystals are mitogenic and induce oncogene expression and matrix metalloproteinase (MMP) synthesis and secretion in human fibroblasts. To date, BCP crystal-elicited signal transduction pathways have not been completely studied. Because protein kinase C (PKC) is known to play an important role in signal transduction, we investigated the participation of this pathway in the BCP crystal induction of MMP-1 and MMP-3 mRNA and protein expressions in human fibroblasts. Using reverse transcription/polymerase chain reaction (RT-PCR) and Northern and Western blotting techniques, we show here that BCP crystal stimulation of MMP-1 and MMP-3 mRNA and protein expressions in human fibroblasts is dependent upon the calcium-dependent PKC signal transduction pathway and that the PKC alpha isozyme is specifically involved in the pathway. We have previously shown that BCP crystal induction of MMP-1 and MMP-3 is also dependent on the p44/42 mitogen-activated protein kinase (p44/42 MAPK) signal transduction pathway. We now show that these two pathways operate independently and seem to complement each other. This leads to our hypothesis that the two pathways initially function independently, ultimately leading to an increase in mitogenesis and MMP synthesis, and may converge downstream of PKC and p44/42 MAPK to mediate BCP crystal-induced cellular responses.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Sequence
  • Calcium Phosphates / chemistry
  • Calcium Phosphates / pharmacology*
  • Cells, Cultured
  • DNA Primers
  • Enzyme Induction
  • Fibroblasts / drug effects*
  • Fibroblasts / enzymology
  • Humans
  • Isoenzymes / metabolism*
  • MAP Kinase Signaling System
  • Matrix Metalloproteinase 1 / biosynthesis*
  • Matrix Metalloproteinase 3 / biosynthesis*
  • Protein Kinase C / metabolism*
  • Protein Kinase C-alpha
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction

Substances

  • Calcium Phosphates
  • DNA Primers
  • Isoenzymes
  • alpha-tricalcium phosphate
  • tetracalcium phosphate
  • calcium phosphate, monobasic, anhydrous
  • calcium phosphate
  • PRKCA protein, human
  • Protein Kinase C
  • Protein Kinase C-alpha
  • Matrix Metalloproteinase 3
  • Matrix Metalloproteinase 1
  • calcium phosphate, dibasic, anhydrous