Production of extracellular matrix‐degrading proteinases by primary cultures of human epithelial ovarian carcinoma cells

• Published in: Cancer Vol. 80; no. 8; pp. 1457 - 1463
• Main Authors: Fishman, David A., Bafetti, Lisa M., Banionis, Saulis, Kearns, Alicia S., Chilukuri, Krishna, Stack, M. Sharon
• Format: Journal Article
• Language: English
• Published: New York John Wiley & Sons, Inc 15.10.1997; Wiley-Liss
• Subjects: Adult; Aged; Biological and medical sciences; Collagenases - biosynthesis; Epithelium - enzymology; extracellular matrix; Extracellular Matrix - metabolism; Female; Female genital diseases; Gelatinases - biosynthesis; Gynecology. Andrology. Obstetrics; Humans; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; matrix metalloproteinases; Medical sciences; Metalloendopeptidases - biosynthesis; Middle Aged; Neoplasm Staging; ovarian carcinoma; Ovarian Neoplasms - enzymology; Ovary; plasminogen activators; Plasminogen Activators - biosynthesis; proteinase; Tumor Cells, Cultured; tumor metastasis; Tumors; Human; Cell culture; u-Plasminogen activator; Carcinoma; Serine endopeptidases; Enzyme; Metalloendopeptidases; Malignant tumor; Gelatinase B; Female genital diseases; Gelatinase A; Ovarian diseases; Peptidases; Ovary; Enzymatic activity; Ascitic fluid; Established cell line; Hydrolases; Female; Extracellular matrix; Ascites
• ISSN: 0008-543X; 1097-0142
• DOI: 10.1002/(SICI)1097-0142(19971015)80:8<1457::AID-CNCR13>3.0.CO;2-4

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 (gelatinase B), by short term primary cultures of epithelial ovarian carcinoma cells derived from primary ovarian tumors, intraperitoneal metastases, or ascites. The presence of these enzymatic activities in samples of ascites was also evaluated. The effect of adhesive substratum on proteinase production was determined. METHODS A coupled spectrophotometric 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 activity was evaluated by gelatin zymography. RESULTS Ascitic fluids from 18 patients contained u‐PA, MMP‐2, and MMP‐9. However, short term primary cultures of cells derived from primary ovarian tumors (OVET), metastatic lesions (OVEM), or ascites (OVEA) produced very low levels of u‐PA. Production of u‐PA by OVET and OVEM cells was regulated by adhesive substratum. Conditioned media from OVET, OVEM, and OVEA cells contained 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 substratum. CONCLUSIONS These results demonstrate that primary cultures of epithelial ovarian carcinoma cells derived from three distinct anatomic 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 significant role in the intraperitoneal invasion of ovarian carcinoma cells. Cancer 1997; 80:1457‐63. © 1997 American Cancer Society. Production of extracellular matrix‐degrading proteinases by short term primary cultures of epithelial ovarian carcinoma cells was analyzed. The cells were derived from primary ovarian tumors, intraperitoneal metastases, and ascites. Matrix metalloproteinase‐2 (MMP‐2, gelatinase A) was the predominant proteinase secreted, with lower levels of MMP‐9 (gelatinase B) and urinary‐type plasminogen activator (u‐PA) detected. This suggested that MMP‐2 may play a significant role in the intraperitoneal invasion of epithelial ovarian carcinoma.