PRODUCTION OF EXTRACELLULAR MATRIX-DEGRADING PROTEINASES BY PRIMARY CULTURES OF HUMAN EPITHELIAL OVARIAN-CARCINOMA CELLS

BACKGROUND. The authors analyzed the secretion of extracellular matrix -degrading proteinases, including urinary-type plasminogen activator ( u-PA), matrix metalloproteinase-2 (MMP-2, gelatinase A), and MMP-9 (ge latinase B), by short term primary cultures of epithelial ovarian carc inoma cells derived from primary ovarian tumors, intraperitoneal metas tases, or ascites. The presence of these enzymatic activities in sampl es of ascites was also evaluated. The effect of adhesive substratum on proteinase production was determined. METHODS. A coupled spectrophoto metric assay was utilized to evaluate the initial rate of plasminogen activation by u-PA in conditioned medium; this involved monitoring the activity of generated plasmin with a colorimetric substrate. MMP acti vity was evaluated by gelatin zymography. RESULTS. Ascitic fluids from 18 patients contained u-PA, MMP-2, and MMP-9. However, short term pri mary cultures of cells derived from primary ovarian tumors (OVET), met astatic lesions (OVEM), or ascites (OVEA) produced very low levels of u-PA. Production of u-PA by OVET and OVEM cells was regulated by adhes ive substratum. Conditioned media from OVET, OVEM, and OVEA cells cont ained high levels of both MMP-2 and MMP-9. MMP-9 levels decreased with increasing passage in culture, whereas MMP-2 activity was maintained. Production of neither MMP-2 nor MMP-9 was regulated by adhesive subst ratum. CONCLUSIONS. These results demonstrate that primary cultures of epithelial ovarian carcinoma cells derived from three distinct anatom ic locations produce MMP-2 and MMP-9, with low level secretion of u-PA . These data suggest that MMPs, particularly MMP-2, may play a signifi cant role in the intraperitoneal invasion of ovarian carcinoma cells. (C) 1997 American Cancer Society.