Inhibition of MT1-MMP proteolytic function and ERK1/2 signalling influences cell migration and invasion through changes in MMP-2 and MMP-9 levels

Abstract

Membrane type-1 matrix metalloproteinase (MT1-MMP, MMP-14) is a unique protease that cleaves extracellular proteins, activates proMMPs, and initiates intracellular signalling. MCF-7 cells are non-invasive and deficient in MT1-MMP, MMP-2, and MMP-9 expression. We created an MCF-7 cell line (C2) that stably produces active MT1-MMP and demonstrated increased ERK1/2 phosphorylation. MAPK inhibition in this cell line showed an inverse relationship in MMP-2 and MMP-9 transcripts where levels of these genes increased and decreased, respectively. Using invasive MDA-MB 231 cells that endogenously produce MT1-MMP and have naturally high pERK levels, we demonstrated the identical inverse relationship between MMP-2 and -9 transcript and protein levels, suggesting that this novel relationship is conserved amongst MT1-MMP positive breast cancer cells. To further analyze the relationship between MMP-2 and -9 levels, we chemically inhibited activation and catalytic activity of MT1-MMP using a furin and MMP inhibitor, respectively, to show that interference with the functions of MT1-MMP induced changes in MMP-2 and 9 transcript levels that were always inverse of each other, and likely mediated by differential transcriptional activity of the NF-κB transcription factor. Furthermore, we analyzed the functional consequences of these expression changes to show MMP, and in particular ERK, inhibition decreased migration and invasion using 2D culture, and inhibits the formation of an invasive phenotype in Matrigel 3D culture. This study demonstrated a novel inverse transcriptional relationship between MMP-2 and -9 levels and MT1-MMP activity that have functional consequences, and also showed that increases in the levels of MMPs does not necessarily correlate with an invasive phenotype.

Anticancer effects of isofraxidin against A549 human lung cancer cells via the EGFR signaling pathway

“…The results of the present study revealed that isofraxidin reduced the expression of p-AKT in A549 cells; therefore, isofraxidin may affect the expression of the Bcl-2 protein family by inhibiting the AKT signaling pathway, leading to the subsequent inhibition of the proliferation of A549 cells and the induction of apoptosis. ERK has been reported to regulate the expression of MMP family-associated proteins, thus affecting cell migration and invasion (29,30). The present study also revealed that isofraxidin reduced the expression of p-ERK in A549 cells, indicating that isofraxidin may inhibit the expression of MMP-2 and MMP-9 proteins by inhibiting the ERK signaling pathway, subsequently inhibiting the migration and invasion of A549 cells.…”

Multiple Biological Effects of an Iridoid Glucoside, Catalpol, and Its Underlying Molecular Mechanisms

“…Patients receiving catalpol had significantly lower serum levels of colon cancer biomarkers of carbohydrate antigen 19-9 (CA 19-9), carcinoembryonic antigen (CEA), MMP-2, and MMP-9 as compared to the placebo group. The reduction in these biomarkers may indicate that catalpol reduced metastases in colon cancer patients [89]. Adverse effects were significantly fewer that in patients receiving bevacizumab and, in contrast to bevacizumab, there were no fatal adverse effects [80].…”

The proprotein convertase furin in tumour progression

“…38 Interestingly, inhibition of furin through expression of its prosegment reduced the invasive phenotype of breast cancer cell lines in vitro, whereas expression of the prosegment of PCSK6 had the inverse effect, pointing at opposing roles of furin and PCSK6 in breast cancer motility and invasiveness. 39 In another in vitro study furin inhibition was used to decrease MMP14 activity in breast cancer cells, which resulted in changed levels of MMP2 and MMP9, 40 indicating a potential link between furin activity, MMP expression and motility in breast cancer cells. Additionally, Scamuffa et al showed that overexpression of the inhibitory prosegment of furin in MDA-MB-231 cells results in impaired processing of the substrates platelet-derived growth factor A (PDGF-A aka PDGF-1) and IGF1R and delayed tumour growth in vivo.…”