Membrane-Type-3 Matrix Metalloproteinase (MT3-MMP) Functions as a Matrix Composition-Dependent Effector of Melanoma Cell Invasion

Abstract

In primary human melanoma, the membrane-type matrix metalloproteinase, MT3-MMP, is overexpressed in the most aggressive nodular-type tumors. Unlike MT1-MMP and MT2-MMP, which promote cell invasion through basement membranes and collagen type I-rich tissues, the function of MT3-MMP in tumor progression remains unclear. Here, we demonstrate that MT3-MMP inhibits MT1-MMP-driven melanoma cell invasion in three-dimensional collagen, while yielding an altered, yet MT1-MMP-dependent, form of expansive growth behavior that phenocopies the formation of nodular cell colonies. In melanoma cell lines originating from advanced primary or metastatic lesions, endogenous MT3-MMP expression was associated with limited collagen-invasive potential. In the cell lines with highest MT3-MMP expression relative to MT1-MMP, collagen-invasive activity was increased following stable MT3-MMP gene silencing. Consistently, MT3-MMP overexpression in cells derived from less advanced superficially spreading melanoma lesions, or in the MT3-MMP knockdown cells, reduced MT1-MMP-dependent collagen invasion. Rather than altering MT1-MMP transcription, MT3-MMP interacted with MT1-MMP in membrane complexes and reduced its cell surface expression. By contrast, as a potent fibrinolytic enzyme, MT3-MMP induced efficient invasion of the cells in fibrin, a provisional matrix component frequently found at tumor-host tissue interfaces and perivascular spaces of melanoma. Since MT3-MMP was significantly upregulated in biopsies of human melanoma metastases, these results identify MT3-MMP as a matrix-dependent modifier of the invasive tumor cell functions during melanoma progression.

MMP16 Mediates a Proteolytic Switch to Promote Cell–Cell Adhesion, Collagen Alignment, and Lymphatic Invasion in Melanoma

“…We reported that MMP16 functions as a matrix compositiondependent regulator of melanoma cell invasion and 3D growth (16). While promoting fibrin invasion, MMP16 also associates with MMP14 in hetero-oligomers cleaving and posttranslationally suppressing this proinvasive protease (16). These observations suggest an MMP16 function in melanoma pathogenesis.…”

“…WM852 and WM165 cells from melanoma metastasis (Wistar Institute, Philadephia, PA), as well as Bowes melanoma cells and COS1 cells (ATCC) were cultivated as described (16,17). Human foreskin lymphatic endothelial cells (LEC; PromoCell), and blood endothelial cells (BEC; dermal HDBECs and umbilical vein endothelial cells HUVECs, PromoCell) were cultured in Endothelial Cell Growth Medium MV containing gentamicin (50 mg/mL).…”

“…Ten spheroids were mixed with 5,000 melanoma cells and 50 mL fibrin (4.5 mg/mL; ref. 16). After 3 to 5 days, the cultures were fixed, subjected to immunofluorescence, and imaged using Leica SP5 confocal microscope.…”

“…Although, for example, collagen III, laminin, fibrin, CD44, and LRP1 are cleaved by both MT-MMPs in vitro, potential in vivo functions of MMP16 in tumor cells have remained unclear (15). We reported that MMP16 functions as a matrix compositiondependent regulator of melanoma cell invasion and 3D growth (16). While promoting fibrin invasion, MMP16 also associates with MMP14 in hetero-oligomers cleaving and posttranslationally suppressing this proinvasive protease (16).…”

Membranous type matrix metalloproteinase 16 induces human prostate cancer metastasis

“…MMP16 was originally cloned from a human placenta cDNA library, and was demonstrated to be located in the cell membrane (13). MMP16 exhibits a high expression level in a variety of tumors tissues, including gastric cancer, astrocytoma and melanoma, compared with normal tissues (14)(15)(16), indicating its potential biological function. A previous study reported that the migration and invasion of gliomas was mediated by MMP16 (17), which indicated that the expression of the MMP16 gene may be associated with tumor cell invasion and metastasis.…”