Elsevier

Gynecologic Oncology

Volume 112, Issue 2, February 2009, Pages 319-324
Gynecologic Oncology

Expression of membrane type 1 matrix metalloproteinase (MMP-14) in epithelial ovarian cancer: High level expression in clear cell carcinoma

https://doi.org/10.1016/j.ygyno.2008.09.025Get rights and content

Abstract

Objective

Clear cell carcinomas of the ovary constitute approximately 5% of all ovarian neoplasms and have a distinct gene expression profile relative to other ovarian carcinoma histotypes. Tumors often present as an early stage large pelvic mass with a high degree of recurrence and frequent early metastasis. Matrix metalloproteinases (MMPs) play a role in intraperitoneal metastasis through breakdown of cell-cell and cell-matrix barriers, enabling anchoring of secondary lesions and promoting proliferation in a geometrically constrained matrix environment. The objective of this study was to evaluate MMP expression in ovarian clear cell carcinoma.

Methods

Immunohistochemistry was used to evaluate expression of membrane type 1 MMP (MMP-14), MMP-2 and MMP-9 in a panel of ovarian tumors. Western blotting and gelatin zymography were used to examine MMP-14 expression and activity in the clear cell carcinoma cell line ES2. The ability of ES2 cells to invade and proliferate within three-dimensional collagen gels was evaluated.

Results

High level expression of MMP-14 and MMP-2 were observed in ovarian clear cell carcinoma relative to other histotypes (94–95% strong positive). MMP-14 was expressed and active in cultured ES2 cells. ES2 cells also exhibited MMP-dependent invasion of and proliferation within three-dimensional collagen gels.

Conclusions

The high level expression of MMP-14 together with in vitro functional analyses suggest that MMP-14 may contribute to both the proliferative capacity and the enhanced parenchymal metastasis of ovarian clear cell carcinoma.

Introduction

Ovarian cancer is the major cause of death from gynecologic malignancy in the United States, resulting in over 15,000 deaths/year [1], as most patients succumb to metastatic disease. Tumors exhibit marked heterogeneity and have been classified into serous, mucinous, endometrioid, and clear cell on the basis of morphology [2], [3], [4]. More recently, large scale gene expression analyses have demonstrated that clear cell carcinomas exhibit a distinct gene expression profile, with upregulation of genes involved in drug detoxification, chemoresistance, and growth signaling [2], [5], [6]. Clear cell carcinomas of the ovary constitute approximately 5% of all ovarian neoplasms. Clinically, clear cell carcinomas often present as a large pelvic mass, the majority of which are detected at an early stage (FIGO stage I) [7], [8], [9]. Despite the early stage diagnosis, survival rates are significantly lower for women with clear cell carcinoma relative to stage-matched serous adenocarcinoma of the ovary [9], [10]. Furthermore, tumors are more chemoresistant [11], resulting in a high degree of recurrence [7] and exhibit more frequent early metastasis to lymph nodes and parenchymal organs [12].

Matrix metalloproteinases (MMPs) have been implicated in a large variety of biologic processes resulting from proteolysis of structural and signaling components of the extracellular matrix and cell surface, thereby influencing differentiation, migration, tissue invasion, and proliferation [reviewed in [13], [14]]]. The membrane-anchored collagenase designated membrane type 1 MMP (MT1-MMP, MMP-14) is overexpressed in malignant ovarian tumors [15], [16], [17], [18], [19], [20] and expression correlates with poor survival [16], [17], [18]. MMP-14 forms a trimolecular complex on the cell surface containing tissue inhibitor of metalloproteinases 2 (TIMP-2) and proMMP-2, resulting in proMMP-2 activation and release [13], [14]. While TIMP-2 levels were reported to be significantly elevated in ovarian clear cell carcinoma [21], the expression of MMP-14 has not been evaluated. In addition to functioning as a proMMP-2 activator, recent studies have demonstrated that MMP-14-catalyzed pericellular collagenolysis promotes tissue invasion of a number of tumor cells [15], [22], [23], [24], [25] and enables cells to defeat growth regulatory checkpoints, thereby enabling proliferation in an otherwise constrained three-dimensional collagen or fibrin microenvironment [26].

In the current study, ovarian clear cell tumors were evaluated using immunohistochemistry for expression of MMP-14, MMP-2 and MMP-9. Expression levels were compared to those in other histotypes (serous, endometrioid, mucinous). Complementary in vitro analyses examined MMP-14 expression and function in the clear cell ovarian carcinoma cell line ES2.

Section snippets

Case selection and tissue microarray

A total of 163 cases of primary epithelial ovarian tumors from oopherectomies performed at Northwestern Memorial Hospital during the period of 1999 to 2003 were collected with Institutional Review Board-approved consent from the Surgical Pathology archives. Paraffin-embedded donor tissue blocks were sampled with 1.5 mm punchers using a Beecher tissue microarray (TMA) instrument (Beecher Instruments Inc, Sun Prarie, WI). Gross and histological diagnoses were confirmed in all cases by two

MMP immunohistochemical evaluation

To assess the potential contribution of MMP-14 and MMP-2 to ovarian clear cell carcinomas, immunohistochemical staining was evaluated by histotype, with the related secreted MMP-9 included as a control. Ninety-five percent of clear cell carcinomas (17/18) expressed moderate to high (2 +–3 +) levels of MMP-14 (Table 1, Fig. 1) compared with 45% (32/70) of serous carcinomas, 52% (23/44) of endometroid carcinomas, and 55% (5/9) of mucinous carcinomas (Table 1). Conversely, only one clear cell

Discussion

Clear cell carcinomas of the ovary comprise approximately 5% of all ovarian neoplasms and display a distinct gene expression profile relative to other histotypes [2], [5], [6]. Ovarian clear cell carcinomas exhibit unique features including a more aggressive clinical course and more malignant behavior relative to stage-matched serous adenocarcinomas [7], [9], [11], [12]. The current immunohistochemical analysis of a panel of ovarian tumors demonstrates markedly strong expression of MMP-14 in

Conflict of interest statement

The authors have no conflicts of interest to declare.

Acknowledgments

The authors acknowledge the Pathology Core Facility, the Biostatistics Core Facility, and Dr. Alfred Rademaker of the Northwestern University Robert H. Lurie Comprehensive Cancer Center for their assistance with this study. This work was supported by research grants RO1CA86984 (M.S.S.) and RO1CA109545 (M.S.S.) from the National Institutes of Health, National Cancer Institute.

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