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Hsp90β and p130cas: novel regulatory factors of MMP-13 expression in human osteoarthritic chondrocytes
  1. Zhiyong Fan
  1. Osteoarthritis Research Unit, University of Montreal Hospital Centre, Notre-Dame Hospital, Canada
    1. Ginette Tardif
    1. Osteoarthritis Research Unit, University of Montreal Hospital Centre, Notre-Dame Hospital, Canada
      1. David Hum
      1. Osteoarthritis Research Unit, University of Montreal Hospital Centre, Notre-Dame Hospital, Canada
        1. Nicolas Duval
        1. Pavillon des Charmilles, Canada
          1. Jean-Pierre Pelletier
          1. Osteoarthritis Research Unit, University of Montreal Hospital Centre, Notre-Dame Hospital, Canada
            1. Johanne Martel-Pelletier (jm{at}martelpelletier.ca)
            1. Osteoarthritis Research Unit, University of Montreal Hospital Centre, Notre-Dame Hospital, Canada

              Abstract

              Background: We previously classified human OA chondrocytes into L (Low)- and H (High)-OA according to MMP-13 basal levels and IL-1β inducibility. In H-OA chondrocytes, the regulatory proteins p130cas and NMP4 acting on the MMP-13 promoter were identified.

              Objective: To identify regulators of MMP-13 expression/production in human L-OA chondrocytes, determine their effect on the expression of other MMPs and the effect of IL-1β on these molecules.

              Methods: The identification of the L-OA chondrocyte proteins interacting specifically with the MMP-13 promoter AGRE-site was performed by mass spectrometry. Hsp90β, p130cas and NMP4 siRNAs were transfected into L-OA chondrocytes and incubated with or without IL-1β. Gene expression was determined by real-time PCR, MMP-1 and -13 production by ELISA, and signaling pathway activation by Western blotting and ELISA.

              Results: Hsp90β was identified as a protein of the L-OA/AGRE specific complex. Silencing p130cas and Hsp90β significantly increased the expression (about 4 and 2 fold, respectively) and production of MMP-13. sip130cas affected to a lesser extent MMP-1 expression (2 fold) and production. siNMP4 showed no effect. MMP-2, -3, -9 and -14 expressions were unaffected. Silencing both Hsp90β and p130cas had a significant additive effect on MMP-13, but not MMP-1 expression, the level of which was similar to sip130cas alone. IL-1β decreased p130cas and Hsp90β expression/production, indicating another pathway by which this cytokine up-regulates MMP expression. The IL-1β-triggered signaling pathways responsible for MMP up-regulation were unaffected in the silenced cells.

              Conclusion: This study illustrates the complex regulation of MMP-13 by showing the inhibitory effect of the two cytoplasmic molecules, p130cas and Hsp90β, in L-OA chondrocytes.

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