Elongation and secretion of tissue inhibitor of metalloproteinases 1 by human microvascular endothelial cells cultured in collagen gels is stimulated by mitomycin c.
During angiogenesis, interactions between endothelial cells (ECs) and the surrounding extracellular matrix are influenced by matrix metalloproteinases (MMPs) and their cognate inhibitors, the TIMPs. The authors discovered that the secretion of TIMP-1 by human microvascular ECs (hmECs) cultured within gels of native, fibrillar collagen was increased robustly by mitomycin C (MMC), an inhibitor of cell proliferation. In contrast, hmECs cultured on plastic coated with gelatin or with native fibrillar collagen exhibited nil (on gelatin) or very modest (on native collagen) increases in TIMP-1 upon exposure to MMC. Notably, none of the cultures altered the secretion of TIMP-2, or MMP-1 and -2, in response to MMC. hmECs cultured within collagen gels elongated significantly after exposure to MMC, a response the authors concluded was mediated by TIMP-1, because elongation could be inhibited completely with a function-blocking antibody to TIMP-1. Moreover, substitution of purified human TIMP-1 for MMC induced a similar elongation by hmECs. hmECs cultured within collagen gels did not proliferate under the conditions used in this study; therefore, inhibited proliferation was not a factor in the altered cell shape and TIMP-1 secretion elicited by MMC. These results illustrate that antiproliferative compounds should be used with caution in studies of MMP regulation by ECs.
Hamner
Margaret A
MA
Division of Gerontology and Geriatric Medicine, Department of Medicine, University of Washington, Seattle, WA 98104-2499, USA.
Vernon
Robert B
RB
Gooden
Michel D
MD
Koike
Teruhiko
T
Reed
May J
MJ
D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D003094:Collagen; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism*; D005782:Gels; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D016685:Mitomycin / Q000494:pharmacology*; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology*; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000378:metabolism*
D005782:Gels; D019384:Nucleic Acid Synthesis Inhibitors; D019715:Tissue Inhibitor of Metalloproteinase-1; D016685:Mitomycin; D003094:Collagen; D020778:Matrix Metalloproteinase 2; D020781:Matrix Metalloproteinase 1