MT1-MMP regulates VEGF-A expression through a complex with VEGFR-2 and Src

Abstract

SummaryMembrane-type-1 matrix metalloproteinase (MT1-MMP) is a zinc-dependent type-I transmembrane metalloproteinase involved in pericellular proteolysis, migration and invasion, with elevated levels correlating with a poor prognosis in cancer. MT1-MMP-mediated transcriptional regulation of genes in cancer cells can contribute to tumour growth, although this is poorly understood at a mechanistic level. In this study, we investigated the mechanism by which MT1-MMP regulates the expression of VEGF-A in breast cancer cells. We discovered that MT1-MMP regulates VEGFR-2 cell surface localisation and forms a complex with VEGFR-2 and Src that is dependent on the MT1-MMP hemopexin domain and independent of its catalytic activity. Although the localisation of VEGFR-2 was independent of the catalytic and intracellular domain of MT1-MMP, intracellular signalling dependent on VEGFR-2 activity leading to VEGF-A transcription still required the MT1-MMP catalytic and intracellular domain, including residues Y573, C574 and DKV582. However, there was redundancy in the function of the catalytic activity of MT1-MMP, as this could be substituted with MMP-2 or MMP-7 in cells expressing inactive MT1-MMP. The signalling cascade dependent on the MT1-MMP-VEGFR-2-Src complex activated Akt and mTOR, ultimately leading to increased VEGF-A transcription.

Inhibition of Matrix Metalloproteinase 14 (MMP-14)-mediated Cancer Cell Migration

“…Expression of MMP-14 in cells results in activation of downstream signaling molecules, including ERK1/2 and AKT, both of which are required for MMP-14-mediated cell migration (37,38). Given the fact that deletion of the cytoplasmic domain of MMP-14 does not affect MMP-14-enhanced cell migration and invasion (36,39), it is likely that intracellular signaling relies on interactions with cell surface proteins (40), particularly CD44, which colocalizes with MMP-14 and has been reported previ- ously to be necessary for the migration-promoting activity (9,33,41).…”

Matrix metalloproteinases in head and neck cancer: current perspectives

“…In addition, Eisenach et al described an MMP14-VEGFR2-Src complex formation that controls VEGFR2 cell-surface localization through hemopexin-dependent activity in breast cancer cells. 121 As a result of this complex formation, Akt and mTOR are activated, leading to enhanced VEGFA transcription. 121 Cross-talk signaling between MMP14 and CD44 has also been proposed for phosphorylation of the EGF receptor, leading to the activation of the MAPK and PI3K signaling cascade and consequent migration of Cos1 cells.…”

“…121 As a result of this complex formation, Akt and mTOR are activated, leading to enhanced VEGFA transcription. 121 Cross-talk signaling between MMP14 and CD44 has also been proposed for phosphorylation of the EGF receptor, leading to the activation of the MAPK and PI3K signaling cascade and consequent migration of Cos1 cells. 122 The MMP3 PEX domain has been found to induce hyperplastic growth in orthotopic transplants of lentivirally transduced mammary epithelial cells, even in the complete absence of its active domain, resulting in a nonproteolytic interaction with the Wnt ligand.…”

Membrane Type 1 Matrix Metalloproteinase (MT1-MMP) Ubiquitination at Lys581 Increases Cellular Invasion through Type I Collagen

“…Immunoprecipitation and Protein Immunoblotting-Immunoprecipitation was performed as described previously (41). Briefly, cells were lysed in immunoprecipitation buffer (10 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1% (v/v) Triton X-100, 0.5% (v/v) Nonidet P-40, 1 mM EDTA, 1 mM EGTA, 1 mM sodium vanadate).…”