Induction of matrix metalloproteinase gene expression in an endothelial cell line by direct interaction with malignant cells

Abstract

Mouse endothelial TKD2 cells in monolayers were cocultured with various human cell lines for 24 h, and the expression of several secreted matrix metalloproteinases (MMP) and cell adhesion molecules was examined by real-time reverse transcription-polymerase chain reaction using mouse-specific primers. Coculture with normal fibroblasts did not elicit the expression of these molecules, but coculture with cancer cells induced the expression of MMP-3, MMP-9 and MMP-10 mRNA in endothelial cells, and in normal mouse embryonic fibroblasts. The induction of MMP mRNA was dependent on direct cell adhesion, as separate culture of A549 cells in Boyden chambers did not induce MMP mRNA, and neutralizing antibody against VLA-4 abolished the induction. An inhibitor of phosphatidylinositol-3-phosphate kinase strongly suppressed the induction of MMP-3, MMP-9 and MMP-10 mRNA, and expression of the dominant-negative mutant of phosphatidylinositol-3-phosphate kinase also decreased the induction. It was suggested that intracellular reactive oxygen species (ROS) levels were increased in TKD2 cells following adhesion to cancer cells. ROS scavengers decreased the levels of MMP induction, and roterone, an inhibitor of mitochondrial complex I, strongly suppressed the induction of MMP-3, MMP-9 and MMP-10. The depletion of mitochondria in TKD2 cells decreased the induction of MMP-9, but the induction of MMP-3 and MMP-10 was not affected. These results indicate that the adhesion of cancer cells to endothelial cells activates several distinct signaling pathways to induce MMP gene expression, and the pathways for MMP-3, MMP-9 and MMP-10 are partly different. For the induction of MMP-9, mitochondria participate in induction, possibly through the production of ROS.

What does matrix metalloproteinase-1 expression in patients with breast cancer really tell us?

“…The specific contributions of the cancer-associated fibroblasts to tumor growth are poorly understood, but it has been suggested that they are able to promote the growth of mammary carcinoma cells and to enhance tumor angiogenesis [28-30], as well as through the secretion of proteinases (including MMP-1) and other proteins (for example, stromal-cell derived factor 1 (also called CXCL 12), syndecan-1, CXCR4 and Caveolin-1) [31,32]. The molecular profile of the lethal breast cancer microenvironment is based on activated cancer-associated fibroblasts and revealed also by high levels of interstitial collagenase MMP-1 (both at protein and mRNA levels), which facilitate angiogenesis and increase ECM degradation, both crucial processes for the invasive and migratory phenotype of metastatic BC [14].…”

T-Cell Production of Matrix Metalloproteinases and Inhibition of Parasite Clearance by TIMP-1 During ChronicToxoplasmaInfection in the Brain

“…To focus on the kinetics of MMP-8, MMP-10 and TIMP-1 observed by array, kinetic analysis was conducted over eight time points (3, 7, 14, 21, 28, 35, 42 and 60 days post-infection) and compared with expression in uninfected brains. Real-time PCR amplification was performed using primer sequences as previously described (Hasebe et al, 2007) for MMP-10, forward (5′-CCTGTGTTGTCTGTCTCTCCA-3′), reverse (5′-CGTGCTGACTGAATCAAAGGA-3′); and designed for MMP-8, forward (5′-ACGGAGTGAGAGGTGTGGAT-3′), reverse (5′-TCTGCCTGGGAACTTATTGG-3′); and TIMP-1, forward (5′-ATCTGGCATCCTCTTGTTGC-3′), reverse (5′-CATTTCCCACAGCCTTGAAT-3′). DNA was amplified using a Bio-Rad iCycler in the presence of SYBR Green.…”

Matrix metalloproteinase-9 involvement in the structural plasticity of dendritic spines

“…The expression of MMP-9 is regulated on multiple levels: (1) transcription (for a comprehensive review of the transcriptional and epigenetic regulation of MMP-9 expression, including regulation through non-coding RNAs, see Labrie and St-Pierre, 2013; several studies also indicate that reactive oxygen species (ROS) activate MMP-9 expression through AP-1 transcription factors; Hasebe et al, 2007; Hsieh et al, 2014); (2) posttranslation (also involving non-proteolytic activation); (3) local translation (Dziembowska et al, 2012); (4) sequestration on the cell membrane (e.g., binding to cell adhesion molecules, such as hyaluronian receptor CD44 (Bourguignon et al, 1998), integrins (Wang et al, 2003), lipoprotein receptor-related protein-1 (LRP-1), and megalin/LRP-2 (Van den Steen et al, 2006)); (5) internalization (Hahn-Dantona et al, 2001); and (6) delayed activation that involves cleavage of the propeptide and co-secretion with TIMP-1, its endogenous inhibitor (Sbai et al, 2010). In the brain, the primary transcriptional regulators of MMP-9 expression are AP-1 and nuclear factor-κB (NF-κB).…”