Elsevier

Oral Oncology

Volume 41, Issue 3, March 2005, Pages 283-293
Oral Oncology

Expression and clinical significance of matrix metalloproteinase-2 and matrix metalloproteinase-9 in oral squamous cell carcinoma

https://doi.org/10.1016/j.oraloncology.2004.08.013Get rights and content

Summary

To successfully establish a metastasis from an invasive carcinoma, the first step involves the degradation of the underlying basement membrane, which is mainly made up of type IV collagen. Matrix metalloproteinases (MMPs)-2 and -9 are thought to play an important role in its degradation because of their ability to destroy this type of collagen.

In order to evaluate the prognostic significance of these proteases, we studied the expression of MMP-2 and -9 in series of 68 OSCC by immunohistochemistry.

Of the oral carcinomas, 28% (n = 19) expressed MMP-2, and 17.6% (n = 12) expressed MMP-9. MMP-2 immunoreactivity was significantly higher in patients with alcohol consumption (p = 0.028) (OR = 4), and in those younger than 60 years (p = 0.041). MMP-9 immunostaining showed statistically significant association with the tumor grade of differentiation (p = 0.019), the T-stage (p = 0.05), and also with the alcohol intake (p = 0.04) (OR = 7.67). In the present study, although not statistically significant, we observed that immunoexpression of MMP-2 and MMP-9 was stronger in patients with lymph node metastasis (OR = 1.65 and 2.29, respectively). In patients without regional lymph node metastasis, positive MMP-9 immunostaining was related to poor survival rates (p = 0.02; OR = 5.8).

MMP-2 and -9 are involved in the invasion process of oral cancer, and MMP-9 is related to poor prognosis in the subset of patients without neck node metastasis. Ethanol could enhance the carcinogenetic process in oral cavity through its influence in the expression of MMP-2 and -9.

Introduction

Oral squamous cell carcinoma (OSCC), the sixth most common cancer, is characterized by a high degree of local invasiveness and a high rate of metastasis to cervical lymph nodes, but a low rate of metastasis to distant organs. Death as a result of cancer is often the result of local recurrence or regional and/or systemic metastasis. Thus, metastasis are a major problem in successful cancer treatment, and it is believed that they begin early in the growth of the primary tumor.1

The process of metastasis consists of series of tumor–host interactions that involve multiple extracellular matrix degrading enzymes, including serine proteinases, cysteine proteinases, and matrix metalloproteinases (MMPs).2 MMPs play an important role in the malignant behavior of the neoplasm. They are a family of Zn2+-dependent proteinases that can degrade all components of extracellular matrix (ECM).3 This family of endopeptidases is associated with ECM degradation in physiological and pathological conditions such as embryonic development, wound healing, angiogenesis, arthritis, inflammation, and tumor metastasis.4 They are classified into four groups according to their substrate specificity: collagenases (MMP-1, -8, and -13), gelatinases (MMP-2 and -9), stromelysins (MMP-3, -10, and -11), and membrane-type MMPs (MT-MMPs) (MMPs 14–17, 24, and 25). Alignment of the amino acid sequences between these enzymes shows that there is a high degree of homology between the enzymes in each group (about 80%) and between groups (about 50%).5 MMPs are produced by cancer cells or through the induction of surrounding stromal cells. This suggests that tumor cells are capable of utilizing MMPs produced by stromal cells and indicates an active role for stroma in tumor invasion.5

To successfully establish a metastasis, the first step is active migration of cancer cells from their tissue of origin and involves the degradation of the underlying basement membrane, which is made up of matrix macromolecules such as type IV collagen, laminin, and heparan sulfate proteoglicans. Thus, this membrane constitutes the first barrier to tumor invasion and because one of its major constituents is type IV collagen, the gelatinases (MMP-2 and MMP-9) are thought to play an important role in its degradation because of their ability to cleave this type of collagen. Most of the MMPs except MT-MMPs and MMP-11 (stromelysin 3) are secreted as inactive zymogens and are activated extracellularly or pericellularly by serine proteinases, such as trypsin, plasmin, and neutrophil elastase6 that cleave their amino-terminal domains. Similarly to other MMPs, MMP-2 is secreted in an inactive zymogen form (pro-MMP2), but its conversion to an active form (active-MMP2) is due to membrane type 1-metalloproteinase (MT1-MMP or MMP-14).7 In fact, pro-MMP-2 binds to TIMP-2 in combination with MT1-MMP on the cell surface, forming a ternary complex. Then, pro-MMP-2 in the complex is activated by adjacent MT1-MMP that is free from TIMP-2.8 Together with the degradation of the basement membrane, cancer expansion requires the induction of angiogenesis in the malignant tissue to provide nourishment for proliferating cancer cells.1 Schnapper et al.9 have demonstrated the importance of type IV collagenases in new blood vessel formation.

Neck lymph node metastasis are found in approximately 50% of patients with OSCC treated by neck dissection,10 and is considered to be the most important clinical prognostic factor in these patients. Although treatment guidelines are based on tumor site, node and distant metastasis (TNM system), they do not always predict clinical outcome accurately, because patients of the same TNM groups have different survival rates. As a consequence, from a prognostic point of view, it is of paramount importance for the clinician to know the invasiveness of the cancer and the likelihood of metastasis. The expressions of MMP family members in OSCC have been reported. Specifically, previous studies have shown that MMP-2 (72-kDa type IV collagenase or gelatinase A)11, 12, 13 and MMP-9 (92-kDa type IV collagenase or gelatinase B)13 are expressed in OSCC. However, the correlation of their expressions with clinical and histopathological features is still controversial.

Studies that elucidate the mechanism of metastasis of cancers may result in the development of treatments which will decrease, or even inhibit this process, improving the survival rate of patients. The purpose of this study was to investigate the expression of MMP-2 and MMP-9 in a series of 68 OSCC, and to evaluate the implications of their expression on clinicopathologic factors, principally on prognosis, using immunohistochemistry.

Section snippets

Patients

This study is based on a retrospective cohort of 68 patients suffering from a primary OSCC who were diagnosed at the Department of Oral and Maxillofacial Surgery, Asturias University Hospital, Oviedo, Spain, between January of 1990 and December of 1992. Inclusion criteria were surgical treatment performed according to standard procedures and consisting of the resection of the primary tumor and a radical or selective ipsi- or bilateral neck dissection, complete clinicopathologic data and

Immunohistologic expression of MMP-2 in primary OSCC and regional lymph node metastasis

Immunoreactivity for MMP-2 was detected in 19 of 68 cases (28%) and primarily in cancer nests of the advancing front of neoplastic tissue (Fig. 1). Of these 19 cases, 3 cases (4.4%) strongly expressed MMP-2; 5 (7.4%) showed immunostaining between 10% and 50% of cancer cells, and finally, 11 (16.2%) expressed less than 10% of positive cell staining. The remaining 49 cases (72%) were negative for MMP-2. Of the 23 cases with lymph node metastasis, 8 cases (34.8%) expressed MMP-2, with 2 (8.7%)

Discussion

Matrix metalloproteinases (MMPs) are a family of proteases commonly expressed in invasive tumors and adjacent stroma, and it is thought that they play an important role in tumor invasion and metastasis. Initially, a carcinoma develops within the epithelium, confined by the basement membrane. This structure has several components (laminin, type IV collagen, and heparan-sulphate proteoglycan) and its degradation is an essential step for progression from carcinoma in situ to invasive carcinoma.

Acknowledgments

The auhors thank Ms. Aurora Fernández García for technical assistance. We are grateful to Dr. Jonas Hannestad (Department of Psychiatry, Duke University) for help in reviewing the manuscript for English syntax. This work was supported by a grant for scientific research from the Ministry of Health, Spain (Instituto de Salud Carlosiii, PI020137).

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