Impaired tyrosine phosphorylation of membrane type 1-matrix metalloproteinase reduces tumor cell proliferation in three-dimensional matrices and abrogates tumor growth in mice

Abstract

Pericellular proteolysis of the extracellular matrix by membrane type 1-matrix metalloproteinase (MT1-MMP) confers tumor cells with the ability to proliferate within three-dimensional (3D) matrices and sustains tumor growth in mice. In this study, we show that in addition to its matrix-degrading activity, phosphorylation of MT1-MMP on its unique tyrosine residue located within its cytoplasmic sequence (Tyr573) may also participate to these processes. Fibrosarcoma cells expressing a proteolytically active but non-phosphorylable mutant of MT1-MMP showed a markedly reduced proliferation rate when embedded within 3D type I collagen matrices, this antiproliferative effect being correlated with arrest in the G(0)/G(1) phase of the cell cycle. Impaired tyrosine phosphorylation of MT1-MMP also inhibits anchorage-independent growth of HT-1080 cells in soft agar as well as their invasion of collagen barriers, two prominent attributes of tumor cells, suggesting a broad inhibitory effect of the MT1-MMP mutant on tumorigenesis. Accordingly, whereas HT-1080 cells formed well-vascularized tumors containing tyrosine-phosphorylated MT1-MMP, tumor growth was completely abolished by expression of the non-phosphorylable MT1-MMP mutant. These findings thus indicate a close co-operation between the matrix-degrading activity of MT1-MMP and tyrosine phosphorylation of its intracellular domain for tumor cell invasion and proliferation and suggest that the targeting of the intracellular signaling pathways leading to tyrosine phosphorylation of MT1-MMP may represent an unexpected alternative strategy for the inhibition of this enzyme.

Oncogenic Src requires a wild-type counterpart to regulate invadopodia maturation

“…MT1-MMP is cited as the key MMP that regulates ECM degradation at invadopodia (Poincloux et al, 2009) and Src kinase activity is known to regulate phosphorylation of MT1-MMP and proteins associated with its trafficking to the cell membrane (Nyalendo et al, 2008;Nyalendo et al, 2007). Future studies should address whether WT Src is necessary to control trafficking to, or activation of MT1-MMP at invadopodia.…”

Epigallocatechin Gallate Targeting of Membrane Type 1 Matrix Metalloproteinase-mediated Src and Janus Kinase/Signal Transducers and Activators of Transcription 3 Signaling Inhibits Transcription of Colony-stimulating Factors 2 and 3 in Mesenchymal Stromal Cells

“…Small interfering RNA and mismatch siRNA were synthesized by Qiagen and annealed to form duplexes. MSCs were transiently transfected with cDNA constructs encoding full-length MT1-MMP (15) or a nonphosphorylatable MT1-MMP mutant Y573F (17).…”

Matrix invasion by tumour cells: a focus on MT1-MMP trafficking to invadopodia

“…It was recently reported that MT1-MMP can be phosphorylated on a tyrosine residue (Tyr573) of the cytoplasmic domain in a manner that is dependent on Src tyrosine kinase, and that this phosphorylation is required for tumour-cell proliferation and invasion of 3D collagen matrices and for tumour growth in nude mice (Nyalendo et al, 2008;Nyalendo et al, 2007). As noted above, the LLY573 sequence is required for clathrin-mediated uptake of MT1-MMP (Uekita et al, 2001), suggesting a possible regulation of endocytosis by phosphorylation downstream of Src.…”