Differential regulation of cytokine-induced MMP-1 and MMP-13 expression by p38 kinase inhibitors in human chondrosarcoma cells: potential role of Runx2 in mediating p38 effects.

Abstract

OBJECTIVE:

To investigate mitogen activated protein (MAP) kinase pathways for their ability to differentially regulate the expression of matrix metalloprotease (MMP)-1 and -13 in human chondrosarcoma cells using pathway-selective inhibitors.

DESIGN:

Human chondrosarcoma cell lines (SW1353 and JJ012) and human articular chondrocytes (HACs) were treated with cytokines (IL-1beta and TNFalpha) and the expression of MMP-1 and -13 was analyzed. The effects of MAP kinase inhibitors on cytokine-induced expression of MMP-1 and -13 were evaluated using ELISA and Western blot analyses. The possible involvement of the Runx2 pathway in mediating p38 effects on MMP-13 expression was analyzed using promoter-reporter assays, ELISA and immunoprecipitation analyses.

RESULTS:

IL-1beta and TNFalpha strongly induced the expression of MMP-1 and -13 in SW1353 cells and HACs, whereas only TNFalpha was found to induce the expression of these two MMPs in JJ012 cells. Cytokine treatment did not result in a significant increase in the activity of MMPs because of the excess production of endogenous tissue inhibitors of metalloproteases (TIMPs). Treatment with p38 kinase inhibitors (SB203580 and SB242235) strongly inhibited cytokine-induced MMP-13 expression in a dose-dependent fashion while having a somewhat weaker inhibitory effect on MMP-1 expression. In contrast, inhibitors of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) pathways did not inhibit the expression of either MMP. Overexpression of Runx2 robustly stimulated the transcriptional activation of MMP-13 but had no effect on MMP-1 expression. Furthermore, IL-1beta induced the phosphorylation of Runx2, and this effect was blocked by a p38 kinase inhibitor. Our data suggest that Runx2 is likely to be a key downstream mediator of p38 effects in the differential regulation of IL-1beta induced MMP-13 expression.

CONCLUSIONS:

These studies demonstrate the differential inhibition of cytokine-induced MMP-1 and -13 expression by p38 kinase inhibitors in human chondrosarcoma cells. Our studies also suggest the involvement of Runx2, at least in part, in mediating the effects of p38 on MMP-13 expression.

• Authors: Yong Pei, Anita Harvey, Xiao-Peng Yu, Srinivasan Chandrasekhar, Kannan Thirunavukkarasu
• Journal: Osteoarthritis and cartilage (Osteoarthritis Cartilage) Vol. 14 Issue 8 Pg. 749-58 (Aug 2006) ISSN: 1063-4584 [Print] England
• PMID: 16549373 (Publication Type: Journal Article)

Chemical References

• Core Binding Factor Alpha 1 Subunit
• Interleukin-1beta
• RUNX2 protein, human
• p38 Mitogen-Activated Protein Kinases
• Matrix Metalloproteinase 13
• Matrix Metalloproteinase 1

Topics

• Blotting, Western (methods)
• Cell Line, Tumor
• Chondrosarcoma (enzymology, immunology)
• Core Binding Factor Alpha 1 Subunit (pharmacology)
• Enzyme-Linked Immunosorbent Assay (methods)
• Humans
• Immunoprecipitation
• Interleukin-1beta (pharmacology)
• Matrix Metalloproteinase 1 (genetics, metabolism)
• Matrix Metalloproteinase 13 (genetics, metabolism)
• Phosphorylation
• Promoter Regions, Genetic
• Reverse Transcriptase Polymerase Chain Reaction
• Transfection (methods)
• p38 Mitogen-Activated Protein Kinases (antagonists & inhibitors)