Molecular and Cellular Endocrinology 187 (2002) 39 – 45 The prognostic value of metalloproteinases and angiogenic factors in ovarian carcinoma Ben Davidson a , Iris Goldberg b , Walter H. Gotlieb c , Juri Kopolovic b , Gilad Ben-Baruch c , Jahn M. Nesland a , Reuven Reich d, * a Department of Pathology, The Norwegian Radium Hospital, Affiliated with the Uniersity of Oslo, Montebello 0310 Oslo, Norway b Department of Pathology, Sheba Medical Center, Affiliated with Sackler School of Medicine, Tel -Hashomer 52621, Israel c Diision of Gynecologic Oncology, Sheba Medical Center, Affiliated with Sackler School of Medicine, Tel -Ai Uniersity, Tel -Hashomer 52621, Israel d Department of Pharmacology, Faculty of Medicine, Hebrew Uniersity, Jerusalem 91120, Israel Abstract The objective of this study was to analyze the correlation between matrix metalloproteinases (MMPs) and angiogenic genes and survival in advanced-stage ovarian carcinomas. Primary and metastatic ovarian carcinomas from patients diagnosed with FIGO stage III–IV disease and followed up to 20 years were studied using mRNA in situ hybridization (ISH). Expression of MMP-2, MMP-9, membrane-type 1-MMP (MT1-MMP), the MMP inhibitor TIMP-2, vascular endothelial growth factor (VEGF), interleukin-8 (IL-8) and basic fibroblast growth factor (bFGF) was studied. MMP-2, MMP-9 and TIMP-2 mRNA was detected in both tumor and stromal cells, while MT1-MMP was largely confined to tumor cells. In univariate analysis of primary tumors, TIMP-2 and MMP-9 mRNA expression correlated with poor outcome. In metastatic lesions, mRNA expression of TIMP-2, MMP-2, and MT1-MMP correlated with poor survival. In a multivariate analysis of primary tumors, TIMP-2 expression in stromal cells (P =0.006) and MMP-9 expression in tumor cells (P =0.011) retained their predictive value. Intense expression of bFGF mRNA and weak expression of IL-8 mRNA was detected in both stromal and tumor cells in most cases, while VEGF mRNA expression was limited to a few cases. Angiogenic mRNA expression showed no correlation with disease outcome in survival analysis (P 0.05). We conclude that bFGF is the major angiogenic factor expressed in ovarian carcinoma at the mRNA level. MMP-2, MMP-9, MT1-MMP and TIMP-2 are valid markers of poor survival in advanced-stage ovarian carcinoma. © 2002 Elsevier Science Ireland Ltd. All rights reserved. Keywords: Ovarian carcinoma; Disease outcome; mRNA in situ hybridization; Matrix metalloproteinases; Angiogenic genes www.elsevier.com/locate/mce 1. Introduction Local invasion, as well as distant spread, of malig- nant neoplasms, involves degradation of subepithelial and -endothelial basement membranes and modification of the extracellular matrix (ECM) (Liotta et al., 1983). This process is now known to depend on complex interactions between tumor cells, stromal cells and the endothelium, mediated by a large number of adhesion molecules, proteolytic enzymes, ECM components and transcription factors (Werb, 1997). Matrix metallo- proteinases (MMPs), a family of zinc- and calcium-de- pendent enzymes, mediate the invasive and metastatic properties of most malignant tumors, in addition to their ubiquitous presence in benign reparative and infl- ammatory processes, through their ability to degrade most ECM components (Crawford and Matrisian, 1994 – 5; Aznavoorian et al., 1993; Liotta and Stetler- Stevenson, 1991). MMP-2 (Gelatinase A, 72 kD type IV collagenase) and MMP-9 (Gelatinase B, 92 kD type IV collagenase) are able to degrade type IV collagen, a component of all basement membranes, thereby facili- tating stromal and vascular invasion by tumor cells (Aznavoorian et al., 1993). The enzymatic activity of MMPs is traditionally postulated to be regulated by tissue inhibitors of metalloproteinases (TIMP), a family presently comprised of four proteins with a molecular size of 21–28 kD. However, recent reports have demon- * Corresponding author. Tel.: +972-2-675-7505; fax: +972-2-675- 8741x8912. E-mail address: reich@yam-suff.cc.huji.ac.il (R. Reich). 0303-7207/02/$ - see front matter © 2002 Elsevier Science Ireland Ltd. All rights reserved. PII:S0303-7207(01)00709-2