Tissue inhibitor of metalloproteinase-3 inhibits neonatal mouse cardiomyocyte proliferation via EGFR/JNK/SP-1 signaling

Am J Physiol Cell Physiol. 2009 Apr;296(4):C735-45. doi: 10.1152/ajpcell.00246.2008. Epub 2009 Feb 11.


We have recently demonstrated that tissue inhibitor of metalloproteinase-3 (TIMP-3) decreases neonatal cardiomyocyte proliferation (Hammoud L, Xiang F, Lu X, Brunner F, Leco K, Feng Q. Cardiovasc Res 75: 359-368, 2007). The aim of the present study was to delineate a pathway through which TIMP-3 exerts its antiproliferative effect. Experiments were conducted on neonatal cardiomyocyte cultures and heart tissues isolated from wild-type (WT) and TIMP-3(-/-) mice. Deficiency in TIMP-3 decreased p27 expression and increased cardiomyocyte proliferation in cardiomyocytes and neonatal hearts. A TIMP-3/epidermal growth factor (EGF) receptor (EGFR)/c-Jun NH(2)-terminal kinase (JNK)/SP-1/p27 pathway was investigated. JNK phosphorylation and EGFR protein levels were increased in TIMP-3(-/-) cardiomyocytes and heart tissues. Treatment with recombinant TIMP-3 decreased JNK phosphorylation and EGFR expression/phosphorylation. Inhibition of JNK activity using SP-600125 decreased SP-1 phosphorylation, increased p27 expression, and decreased cardiomyocyte proliferation. Furthermore, treatment with the EGFR specific inhibitor PD-168393 or the EGF-neutralizing antibody decreased cardiomyocyte proliferation as well as phosphorylation of JNK and SP-1 in both WT and TIMP-3(-/-) cardiomyocytes. We conclude that TIMP-3 inhibits neonatal mouse cardiomyocyte proliferation by upregulating p27 expression. The effects of TIMP-3 are mediated via inhibition of EGFR expression/phosphorylation, and decreases in JNK and SP-1 signaling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Anthracenes / pharmacology
  • Cell Proliferation* / drug effects
  • Cells, Cultured
  • Cyclin-Dependent Kinase Inhibitor p27 / metabolism
  • Epidermal Growth Factor / metabolism
  • ErbB Receptors / antagonists & inhibitors
  • ErbB Receptors / metabolism*
  • JNK Mitogen-Activated Protein Kinases / antagonists & inhibitors
  • JNK Mitogen-Activated Protein Kinases / metabolism*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Myocytes, Cardiac / drug effects
  • Myocytes, Cardiac / enzymology*
  • Phosphorylation
  • Protein Kinase Inhibitors / pharmacology
  • Quinazolines / pharmacology
  • Recombinant Proteins / metabolism
  • Signal Transduction* / drug effects
  • Sp1 Transcription Factor / metabolism*
  • Tissue Inhibitor of Metalloproteinase-3 / deficiency
  • Tissue Inhibitor of Metalloproteinase-3 / genetics
  • Tissue Inhibitor of Metalloproteinase-3 / metabolism*


  • Anthracenes
  • Cdkn1b protein, mouse
  • PD168393
  • Protein Kinase Inhibitors
  • Quinazolines
  • Recombinant Proteins
  • Sp1 Transcription Factor
  • Tissue Inhibitor of Metalloproteinase-3
  • Cyclin-Dependent Kinase Inhibitor p27
  • pyrazolanthrone
  • Epidermal Growth Factor
  • ErbB Receptors
  • JNK Mitogen-Activated Protein Kinases