Induction of tissue inhibitor of metalloproteinases-3 is a delayed early cellular response to hepatocyte growth factor.
Hepatocyte growth factor (HGF) stimulates mitogenic, motogenic, and morphogenic responses in various cell types. We analysed HGF-responsive cells by differential display PCR to identify HGF-induced genes that mediate these biological events. One of the genes identified encoded a member of the tissue inhibitor of metalloproteinases (TIMP) family, TIMP-3. HGF transiently induced TIMP-3 mRNA in keratinocytes as well as kidney and mammary epithelial cells maximally between 4 and 6 h post-stimulation. Increased TIMP-3 protein secretion returned to basal levels within 18 h, while the expression of gelatinases A and B remained unchanged, suggesting that temporary suppression of matrix degradation is a delayed early response to HGF. Ectopic overexpression of TIMP-3 in cultured leiomyosarcoma cells conferred an epithelial morphology, reduced cell growth rate, anchorage-independent growth, and matrix invasion in vitro. Antisense suppression of TIMP-3 was associated with a scattered, fibroblastic cell morphology, as well as enhanced proliferation, anchorage-independent growth, and matrix invasion. A survey of tumor cell lines revealed an inverse relationship between metastatic potential and TIMP-3 expression level. These data suggest that early, transient TIMP-3 expression mediates specific HGF-induced phenotypic changes, and that loss of TIMP-3 expression may enhance the invasion potential of certain tumors.
Laboratory of Cellular and Molecular Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.
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P
Castagnino
J V
JV
Soriano
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R
Montesano
D P
DP
Bottaro
D016475:3T3 Cells; D000818:Animals; D002455:Cell Division; D002460:Cell Line; D048429:Cell Size; D004285:Dogs; D018093:Gelatinases / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D006801:Humans; D051379:Mice; D011994:Recombinant Proteins / Q000494:pharmacology; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D011994:Recombinant Proteins; D019717:Tissue Inhibitor of Metalloproteinase-3; D017228:Hepatocyte Growth Factor; D018093:Gelatinases