INDUCTION OF TISSUE INHIBITOR AND MATRIX METALLOPROTEINASE BY SERUM IN HUMAN HEART-DERIVED FIBROBLAST AND ENDOMYOCARDIAL ENDOTHELIAL-CELLS

To understand the regulatory mechanisms of extracellular matrix (ECM) turnover and proteinase expression in human cardiovascular tissue, we have isolated and characterized human heart fibroblast (HHF) and human heart endothelial (HHE) cells from endomyocardial biopsy specimens. HHE cell in culture exhibited the typical cobblestone growth pattern and positive immunofluorescent staining for factor VIII related antigen. HHF demonstrated the typical spindle shape during culture and were positive for vimentin. Both cell types were negative for alpha-actin, indicating that these eel Is were of nonmuscle origin. Cell growth studies revealed significant growth when maintained in limiting serum concentration, suggesting mitogenic activity of these cells, and demonstrated growth inhibitory activity when grown in serum-free medium. Serum-dependent matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMPs) expression was measured by zymography, immunoblot, and Northern blot analysis. Results indicated that serum induces both the MMP and TIMP expression at the mRNA and protein levels in a dose-dependent manner. This induction was inhibited by actinomycin D and cycloheximide, suggesting transcriptional and translational regulation of MMP and TIMP. Indirect immunofluorescence labeling indicated expression of MMP and TIMP in HHF and HHE cells. These results suggested that the serum induces proliferation as well as expression of MMP and TIMP in HHE and HHF cells. The growth inhibitory activity of these cell cultures will enable us to explore further the nature of this response and compare this phenomenon with other growth inhibitors and growth promoters identified in other normal and transformed cells. (C) 1995 Wiley-Liss, Inc.