Matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 expression and synthetic matrix metalloproteinase-2 inhibitor binding in ovarian carcinomas and tumor cell lines

Saliha Afzal, El Nasir Lalani, William D. Foulkes, Byron Boyce, Simon Tickle, Maria Rosaria Cardillo, Terry Baker, Massimo Pignatelli, Gordon W H Stamp

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

Enhanced matrix metalloproteinase-2 (MMP-2/72-kd type IV collagenase) action correlates with invasion in neoplasia. MMP-2 is inhibited in vivo by tissue inhibitors of metalloproteinases (TIMPs)-TIMP-1 and, especially, TIMP- 2. A synthetic, biotinylated inhibitor specific for activated MMP-2 in solution phase, and immunohistochemistry were used to detect MMP-2 and TIMP- 2 expression in cell lines and ovarian tumors and to analyze the surface- binding capacity of the inhibitors, which are potential therapeutic agents. Characterization of novel monoclonal antibodies to MMP-2 and TIMP-2 is described together with immunocytochemical staining of 83 paraffin-embedded ovarian tumors (67 malignant, 7 borderline, 9 benign) and 9 cell lines. Synthetic MMP-2 inhibitor binding under controlled conditions was visualized by immunofluorescence and avidin-biotin complex immunoperoxidase methods in cell lines and cryostat sections of ovarian tumors. MMP-2 and TIMP-2 showed heterogenous immunoreactivity, with enhanced staining on high-grade tumors, specifically at the invasive front and in vascular invasion. TIMP-2 immunoreactivity was maximal in malignant cell cytoplasm and less intense in desmoplastic fibroblasts. One monoclonal antibody to MMP-2 showed membrane immunoreactivity, apically polarized in benign and low-grade tumors but depolarized and strong in 37 of 44 cases of high-grade invasive tumors. Eleven of eighteen ovarian carcinomas and six of nine cell lines showed membrane localization of the synthetic inhibitor. Maximal binding occurred in the ovarian cell line OVCA 432 and the breast cell lines MCF 7 and MDA MB 435, all of which were immunoreactive for MMP-2. Cell lines propagated on type I collagen showed no enhancement in inhibitor binding. This study demonstrates cell surface binding of a synthetic MMP-2 inhibitor and provides new evidence of MMP-2 and TIMP-2 immunoreactivity in ovarian carcinomas and cell lines.

Original languageEnglish
Pages (from-to)406-421
Number of pages16
JournalLaboratory Investigation
Volume74
Issue number2
Publication statusPublished - Feb 1996
Externally publishedYes

Fingerprint

Tissue Inhibitor of Metalloproteinase-2
Matrix Metalloproteinase Inhibitors
Matrix Metalloproteinase 2
Tumor Cell Line
Matrix Metalloproteinases
Carcinoma
Cell Line
Neoplasms
Monoclonal Antibodies
Staining and Labeling
Tissue Inhibitor of Metalloproteinases
Tissue Inhibitor of Metalloproteinase-1
Membranes
Avidin
Collagenases
Biotin
Collagen Type I
Paraffin
Fluorescent Antibody Technique
Blood Vessels

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 expression and synthetic matrix metalloproteinase-2 inhibitor binding in ovarian carcinomas and tumor cell lines. / Afzal, Saliha; Lalani, El Nasir; Foulkes, William D.; Boyce, Byron; Tickle, Simon; Cardillo, Maria Rosaria; Baker, Terry; Pignatelli, Massimo; Stamp, Gordon W H.

In: Laboratory Investigation, Vol. 74, No. 2, 02.1996, p. 406-421.

Research output: Contribution to journalArticle

Afzal, Saliha ; Lalani, El Nasir ; Foulkes, William D. ; Boyce, Byron ; Tickle, Simon ; Cardillo, Maria Rosaria ; Baker, Terry ; Pignatelli, Massimo ; Stamp, Gordon W H. / Matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-2 expression and synthetic matrix metalloproteinase-2 inhibitor binding in ovarian carcinomas and tumor cell lines. In: Laboratory Investigation. 1996 ; Vol. 74, No. 2. pp. 406-421.
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