Abstract
Pleural malignant mesothelioma is a locally aggressive tumor of mesothelial cell origin. In other tumor types high expression of matrix metalloproteinase (MMP)-2, together with membrane-type1-MMP (MT1-MMP), and low levels of the tissue inhibitor of MMP (TIMP)-2 have been correlated with aggressive tumor progression and low survival rates. Therefore, we compared the expression and activation of these three factors and their regulation by two mesothelioma associated growth factors, platelet-derived growth factor (PDGF)-BB, and transforming growth factor (TGF)-beta1 in six human mesothelioma and one mesothelial cell line. Polymerase chain reaction (PCR), immunoblotting, zymography, and small inhibitory RNAs (siRNA) were used to study gene expression, protein activation, and signal transduction. To proof the relevance of our in vitro data immunohistochemistry was performed in tissue sections. PDGF-BB induced, while TGF-beta1 inhibited cell proliferation. PDGF-BB was a chemoattractant for mesothelial cells, and its effect was increased in the presence of TGF-beta1. TGF-beta1 stimulated the de novo synthesis of pro-MMP-2 in both cell types. Pro-MMP-2 synthesis involved p38 MAP kinase. In cell culture and tissue sections only mesothelial cells expressed MT1-MMP. Migration of mesothelioma cells was dependent on the presence of MT1-MMP. Migration, but not proliferation of mesothelioma cells was inhibited by oleoyl-N-hydroxylamide, TIMP-2, and siRNA for MT1-MMP. Our data suggest that in mesothelioma cells the phosphorylation of p38 MAP kinase is deregulated and is involved in pro-MMP-2 expression. Mesothelioma progression depends on an interaction with mesothelial cells that provide MT1-MMP necessary to activate pro-MMP-2 to facilitate migration through an extracellular matrix (ECM) layer.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Becaplermin
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Cell Line
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Cell Line, Tumor
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Cell Movement / drug effects
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Cell Movement / physiology*
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Cell Proliferation / drug effects
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Epithelial Cells / chemistry
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Epithelial Cells / drug effects
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Epithelial Cells / metabolism
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Humans
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Matrix Metalloproteinase 2 / genetics
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Matrix Metalloproteinase 2 / metabolism
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Matrix Metalloproteinase 9 / genetics
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Matrix Metalloproteinase 9 / metabolism
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Matrix Metalloproteinase Inhibitors
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Matrix Metalloproteinases / genetics
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Matrix Metalloproteinases / metabolism*
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Matrix Metalloproteinases, Membrane-Associated
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Mesothelioma / chemistry
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Mesothelioma / metabolism
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Mesothelioma / pathology
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Mitogen-Activated Protein Kinase 1 / antagonists & inhibitors
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Mitogen-Activated Protein Kinase 1 / genetics
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Mitogen-Activated Protein Kinase 1 / metabolism
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Mitogen-Activated Protein Kinase 3 / antagonists & inhibitors
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Mitogen-Activated Protein Kinase 3 / genetics
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Mitogen-Activated Protein Kinase 3 / metabolism
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Mitogen-Activated Protein Kinases / antagonists & inhibitors
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Mitogen-Activated Protein Kinases / genetics
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Mitogen-Activated Protein Kinases / metabolism*
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Platelet-Derived Growth Factor / pharmacology
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Protease Inhibitors / pharmacology
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Protein Kinase Inhibitors / pharmacology
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Proto-Oncogene Proteins c-sis
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RNA, Small Interfering / genetics
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Tissue Inhibitor of Metalloproteinase-2 / genetics
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Tissue Inhibitor of Metalloproteinase-2 / metabolism
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Tissue Inhibitor of Metalloproteinases / metabolism*
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Transforming Growth Factor beta / pharmacology
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Transforming Growth Factor beta1
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p38 Mitogen-Activated Protein Kinases / antagonists & inhibitors
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p38 Mitogen-Activated Protein Kinases / genetics
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p38 Mitogen-Activated Protein Kinases / metabolism
Substances
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Matrix Metalloproteinase Inhibitors
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Platelet-Derived Growth Factor
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Protease Inhibitors
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Protein Kinase Inhibitors
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Proto-Oncogene Proteins c-sis
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RNA, Small Interfering
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TGFB1 protein, human
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Tissue Inhibitor of Metalloproteinases
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Transforming Growth Factor beta
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Transforming Growth Factor beta1
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Tissue Inhibitor of Metalloproteinase-2
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Becaplermin
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Mitogen-Activated Protein Kinase 1
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Mitogen-Activated Protein Kinase 3
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Mitogen-Activated Protein Kinases
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p38 Mitogen-Activated Protein Kinases
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Matrix Metalloproteinases
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Matrix Metalloproteinases, Membrane-Associated
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Matrix Metalloproteinase 2
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Matrix Metalloproteinase 9