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Molecular Cancer
Published in: Molecular Cancer 1/2016

Open Access 01-12-2016 | Research

Less is more: low expression of MT1-MMP is optimal to promote migration and tumourigenesis of breast cancer cells

Authors: Mario A. Cepeda, Jacob J. H. Pelling, Caitlin L. Evered, Karla C. Williams, Zoey Freedman, Ioana Stan, Jessica A. Willson, Hon S. Leong, Sashko Damjanovski

Published in: Molecular Cancer | Issue 1/2016

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Abstract

Background

Membrane Type-1 Matrix Metalloproteinase (MT1-MMP) is a multifunctional protease implicated in metastatic progression ostensibly due to its ability to degrade extracellular matrix (ECM) components and allow migration of cells through the basement membrane. Despite in vitro studies demonstrating this principle, this knowledge has not translated into the use of MMP inhibitors (MMPi) as effective cancer therapeutics, or been corroborated by evidence of in vivo ECM degradation mediated by MT1-MMP, suggesting that our understanding of the role of MT1-MMP in cancer progression is incomplete.

Methods

MCF-7 and MDA-MB 231 breast cancer cell lines were created that stably overexpress different levels of MT1-MMP. Using 2D culture, we analyzed proMMP-2 activation (gelatin zymography), ECM degradation (fluorescent gelatin), ERK signaling (immunoblot), cell migration (transwell/scratch closure/time-lapse imaging), and viability (colorimetric substrate) to assess how different MT1-MMP levels affect these cellular parameters. We also utilized Matrigel 3D cell culture and avian embryos to examine how different levels of MT1-MMP expression affect morphological changes in 3D culture, and tumourigenecity and extravasation efficiency in vivo.

Results

In 2D culture, breast cancer cells expressing high levels of MT1-MMP were capable of widespread ECM degradation and TIMP-2-mediated proMMP-2 activation, but were not the most migratory. Instead, cells expressing low levels of MT1-MMP were the most migratory, and demonstrated increased viability and ERK activation. In 3D culture, MCF-7 breast cancer cells expressing low levels of MT1-MMP demonstrated an invasive protrusive phenotype, whereas cells expressing high levels of MT1-MMP demonstrated loss of colony structure and cell fragment release. Similarly, in vivo analysis demonstrated increased tumourigenecity and metastatic capability for cells expressing low levels of MT1-MMP, whereas cells expressing high levels were devoid of these qualities despite the production of functional MT1-MMP protein.

Conclusions

This study demonstrates that excessive ECM degradation mediated by high levels of MT1-MMP is not associated with cell migration and tumourigenesis, while low levels of MT1-MMP promote invasion and vascularization in vivo.
Appendix
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Literature
1.
Chambers AF, Groom AC, MacDonald IC. Dissemination and growth of cancer cells in metastatic sites. Nat Rev Cancer. 2002;2(8):563–72.PubMedCrossRef
2.
3.
4.
Leong HS, et al. Invadopodia are required for cancer cell extravasation and are a therapeutic target for metastasis. Cell Rep. 2014;8(5):1558–70.PubMedCrossRef
5.
Visse R, Nagase H. Matrix metalloproteinases and tissue inhibitors of metalloproteinases: structure, function, and biochemistry. Circ Res. 2003;92(8):827–39.PubMedCrossRef
6.
7.
Szabova L, et al. MT1-MMP is required for efficient tumor dissemination in experimental metastatic disease. Oncogene. 2008;27(23):3274–81.PubMedCrossRef
8.
Rowe RG, Weiss SJ. Breaching the basement membrane: who, when and how? Trends Cell Biol. 2008;18(11):560–74.PubMedCrossRef
9.
10.
Chambers AF, Matrisian LM. Changing views of the role of matrix metalloproteinases in metastasis. J Natl Cancer Inst. 1997;89(17):1260–70.PubMedCrossRef
11.
Bourboulia D, Stetler-Stevenson WG. Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs): Positive and negative regulators in tumor cell adhesion. Semin Cancer Biol. 2010;20(3):161–8.PubMedPubMedCentralCrossRef
12.
Egeblad M, Werb Z. New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer. 2002;2(3):161–74.PubMedCrossRef
13.
Eguchi T, et al. Novel transcription-factor-like function of human matrix metalloproteinase 3 regulating the CTGF/CCN2 gene. Mol Cell Biol. 2008;28(7):2391–413.PubMedPubMedCentralCrossRef
14.
Kwan JA, et al. Matrix metalloproteinase-2 (MMP-2) is present in the nucleus of cardiac myocytes and is capable of cleaving poly (ADP-ribose) polymerase (PARP) in vitro. FASEB J. 2004;18(6):690–2.PubMed
15.
Marchant DJ, et al. A new transcriptional role for matrix metalloproteinase-12 in antiviral immunity. Nat Med. 2014;20(5):493–502.PubMedCrossRef
16.
Strongin AY. Proteolytic and non-proteolytic roles of membrane type-1 matrix metalloproteinase in malignancy. Biochim Biophys Acta. 2010;1803(1):133–41.PubMedCrossRef
17.
Seiki M. Membrane-type 1 matrix metalloproteinase: a key enzyme for tumor invasion. Cancer Lett. 2003;194(1):1–11.PubMedCrossRef
18.
Kukreja M, et al. High-throughput multiplexed peptide-centric profiling illustrates both substrate cleavage redundancy and specificity in the MMP family. Chem Biol. 2015;22(8):1122–33.PubMedPubMedCentralCrossRef
19.
Toth M, et al. Pro-MMP-9 activation by the MT1-MMP/MMP-2 axis and MMP-3: role of TIMP-2 and plasma membranes. Biochem Biophys Res Commun. 2003;308(2):386–95.PubMedCrossRef
20.
Itoh Y, et al. Homophilic complex formation of MT1-MMP facilitates proMMP-2 activation on the cell surface and promotes tumor cell invasion. EMBO J. 2001;20(17):4782–93.PubMedPubMedCentralCrossRef
21.
Valacca C, Tassone E, Mignatti P. TIMP-2 interaction with MT1-MMP activates the AKT pathway and protects tumor cells from apoptosis. PLoS One. 2015;10(9):e0136797.PubMedPubMedCentralCrossRef
22.
Gingras D, et al. Activation of the extracellular signal-regulated protein kinase (ERK) cascade by membrane-type-1 matrix metalloproteinase (MT1-MMP). FEBS Lett. 2001;507(2):231–6.PubMedCrossRef
23.
Sakamoto T, Niiya D, Seiki M. Targeting the Warburg effect that arises in tumor cells expressing membrane type-1 matrix metalloproteinase. J Biol Chem. 2011;286(16):14691–704.PubMedPubMedCentralCrossRef
24.
D’Alessio S, et al. Tissue inhibitor of metalloproteinases-2 binding to membrane-type 1 matrix metalloproteinase induces MAPK activation and cell growth by a non-proteolytic mechanism. J Biol Chem. 2008;283(1):87–99.PubMedCrossRef
25.
Sounni NE, et al. Timp-2 binding with cellular MT1-MMP stimulates invasion-promoting MEK/ERK signaling in cancer cells. Int J Cancer. 2010;126(5):1067–78.PubMedPubMedCentral
26.
Mori H, et al. CD44 directs membrane-type 1 matrix metalloproteinase to lamellipodia by associating with its hemopexin-like domain. EMBO J. 2002;21(15):3949–59.PubMedPubMedCentralCrossRef
27.
28.
Rozanov DV, et al. Mutation analysis of membrane type-1 matrix metalloproteinase (MT1-MMP). The role of the cytoplasmic tail Cys(574), the active site Glu(240), and furin cleavage motifs in oligomerization, processing, and self-proteolysis of MT1-MMP expressed in breast carcinoma cells. J Biol Chem. 2001;276(28):25705–14.PubMedCrossRef
29.
Madsen DH, Bugge TH. The source of matrix-degrading enzymes in human cancer: problems of research reproducibility and possible solutions. J Cell Biol. 2015;209(2):195–8.PubMedPubMedCentralCrossRef
30.
Perentes JY, et al. Cancer cell-associated MT1-MMP promotes blood vessel invasion and distant metastasis in triple-negative mammary tumors. Cancer Res. 2011;71(13):4527–38.PubMedCrossRef
31.
Yamamoto H, et al. Overexpression of MT1-MMP is insufficient to increase experimental liver metastasis of human colon cancer cells. Int J Mol Med. 2008;22(6):757–61.PubMed
32.
33.
Figueira RC, et al. Correlation between MMPs and their inhibitors in breast cancer tumor tissue specimens and in cell lines with different metastatic potential. BMC Cancer. 2009;9:20.PubMedPubMedCentralCrossRef
34.
Kohrmann A, et al. Expression of matrix metalloproteinases (MMPs) in primary human breast cancer and breast cancer cell lines: new findings and review of the literature. BMC Cancer. 2009;9:188.PubMedPubMedCentralCrossRef
35.
Artym VV, et al. Dynamic interactions of cortactin and membrane type 1 matrix metalloproteinase at invadopodia: defining the stages of invadopodia formation and function. Cancer Res. 2006;66(6):3034–43.PubMedCrossRef
36.
Nelson CM, Bissell MJ. Modeling dynamic reciprocity: engineering three-dimensional culture models of breast architecture, function, and neoplastic transformation. Semin Cancer Biol. 2005;15(5):342–52.PubMedPubMedCentralCrossRef
37.
Kim Y, et al. Quantification of cancer cell extravasation in vivo. Nat Protoc. 2016;11(5):937–48.PubMedCrossRef
38.
Walsh LA, Cepeda MA, Damjanovski S. Analysis of the MMP-dependent and independent functions of tissue inhibitor of metalloproteinase-2 on the invasiveness of breast cancer cells. J Cell Commun Signal. 2012;6(2):87–95.
39.
Souter LH, et al. Human 21 T breast epithelial cell lines mimic breast cancer progression in vivo and in vitro and show stage-specific gene expression patterns. Lab Invest. 2010;90(8):1247–58.PubMedCrossRef
40.
Hawkes SP, Li H, Taniguchi GT. Zymography and reverse zymography for detecting MMPs and TIMPs. Methods Mol Biol. 2010;622:257–69.PubMedCrossRef
41.
42.
Martin KH, et al. Quantitative measurement of invadopodia-mediated extracellular matrix proteolysis in single and multicellular contexts. J Vis Exp. 2012;66:e4119.PubMed
43.
Cvetkovic D, Goertzen CG, Bhattacharya M. Quantification of breast cancer cell invasiveness using a three-dimensional (3D) model. J Vis Exp. 2014;88.
44.
Barry DJ, et al. Open source software for quantification of cell migration, protrusions, and fluorescence intensities. J Cell Biol. 2015;209(1):163–80.PubMedPubMedCentralCrossRef
45.
Tran TA, et al. Fibroblast Growth Factor Receptor-Dependent and -Independent Paracrine Signaling by Sunitinib-Resistant Renal Cell Carcinoma. Mol Cell Biol. 2016;36(13):1836–55.
46.
Wingfield PT, et al. Biophysical and functional characterization of full-length, recombinant human tissue inhibitor of metalloproteinases-2 (TIMP-2) produced in Escherichia coli. Comparison of wild type and amino-terminal alanine appended variant with implications for the mechanism of TIMP functions. J Biol Chem. 1999;274(30):21362–8.PubMedCrossRef
47.
Stetler-Stevenson WG, Seo DW. TIMP-2: an endogenous inhibitor of angiogenesis. Trends Mol Med. 2005;11(3):97–103.PubMedCrossRef
48.
Worley JR, et al. Sequence motifs of tissue inhibitor of metalloproteinases 2 (TIMP-2) determining progelatinase A (proMMP-2) binding and activation by membrane-type metalloproteinase 1 (MT1-MMP). Biochem J. 2003;372(Pt 3):799–809.PubMedPubMedCentralCrossRef
49.
Maquoi E, et al. Type IV collagen induces matrix metalloproteinase 2 activation in HT1080 fibrosarcoma cells. Exp Cell Res. 2000;261(2):348–59.PubMedCrossRef
50.
51.
Weigelt B, Bissell MJ. Unraveling the microenvironmental influences on the normal mammary gland and breast cancer. Semin Cancer Biol. 2008;18(5):311–21.PubMedPubMedCentralCrossRef
52.
Ribatti D. The chick embryo chorioallantoic membrane as a model for tumor biology. Exp Cell Res. 2014;328(2):314–24.PubMedCrossRef
53.
Band V, et al. Tumor progression in four mammary epithelial cell lines derived from the same patient. Cancer Res. 1990;50(22):7351–7.PubMed
54.
Sounni NE, Noel A. Membrane type-matrix metalloproteinases and tumor progression. Biochimie. 2005;87(3–4):329–42.PubMedCrossRef
55.
Williams KC, Coppolino MG. Phosphorylation of membrane type 1-matrix metalloproteinase (MT1-MMP) and its vesicle-associated membrane protein 7 (VAMP7)-dependent trafficking facilitate cell invasion and migration. J Biol Chem. 2011;286(50):43405–16.PubMedPubMedCentralCrossRef
56.
Tassone E, Valacca C, Mignatti P. Membrane-type 1 matrix metalloproteinase downregulates fibroblast growth factor-2 binding to the cell surface and intracellular signaling. J Cell Physiol. 2015;230(2):366–77.PubMedPubMedCentralCrossRef
57.
Sakamoto T, Seiki M. A membrane protease regulates energy production in macrophages by activating hypoxia-inducible factor-1 via a non-proteolytic mechanism. J Biol Chem. 2010;285(39):29951–64.PubMedPubMedCentralCrossRef
58.
Bramhall SR, et al. Marimastat as maintenance therapy for patients with advanced gastric cancer: a randomised trial. Br J Cancer. 2002;86(12):1864–70.PubMedPubMedCentralCrossRef
59.
Pavlaki M, Zucker S. Matrix metalloproteinase inhibitors (MMPIs): the beginning of phase I or the termination of phase III clinical trials. Cancer Metastasis Rev. 2003;22(2–3):177–203.PubMedCrossRef
60.
Coussens LM, Fingleton B, Matrisian LM. Matrix metalloproteinase inhibitors and cancer: trials and tribulations. Science. 2002;295(5564):2387–92.PubMedCrossRef
61.
62.
Alvarez OA, Carmichael DF, DeClerck YA. Inhibition of collagenolytic activity and metastasis of tumor cells by a recombinant human tissue inhibitor of metalloproteinases. J Natl Cancer Inst. 1990;82(7):589–95.PubMedCrossRef
63.
64.
Sounni NE, et al. Membrane type-1 matrix metalloproteinase and TIMP-2 in tumor angiogenesis. Matrix Biol. 2003;22(1):55–61.PubMedCrossRef
65.
Stetler-Stevenson WG. The tumor microenvironment: regulation by MMP-independent effects of tissue inhibitor of metalloproteinases-2. Cancer Metastasis Rev. 2008;27(1):57–66.PubMedPubMedCentralCrossRef
66.
Wood M, et al. In situ hybridization studies of metalloproteinases 2 and 9 and TIMP-1 and TIMP-2 expression in human prostate cancer. Clin Exp Metastasis. 1997;15(3):246–58.PubMedCrossRef
67.
Visscher DW, et al. Enhanced expression of tissue inhibitor of metalloproteinase-2 (TIMP-2) in the stroma of breast carcinomas correlates with tumor recurrence. Int J Cancer. 1994;59(3):339–44.PubMedCrossRef
68.
Remacle A, et al. High levels of TIMP-2 correlate with adverse prognosis in breast cancer. Int J Cancer. 2000;89(2):118–21.PubMedCrossRef
69.
Ree AH, et al. High levels of messenger RNAs for tissue inhibitors of metalloproteinases (TIMP-1 and TIMP-2) in primary breast carcinomas are associated with development of distant metastases. Clin Cancer Res. 1997;3(9):1623–8.PubMed
70.
Davidson B, et al. MMP-2 and TIMP-2 expression correlates with poor prognosis in cervical carcinoma--a clinicopathologic study using immunohistochemistry and mRNA in situ hybridization. Gynecol Oncol. 1999;73(3):372–82.PubMedCrossRef
71.
Davidson B, et al. High levels of MMP-2, MMP-9, MT1-MMP and TIMP-2 mRNA correlate with poor survival in ovarian carcinoma. Clin Exp Metastasis. 1999;17(10):799–808.PubMedCrossRef
72.
Bradbury A, Pluckthun A. Reproducibility: standardize antibodies used in research. Nature. 2015;518(7537):27–9.PubMedCrossRef
73.
74.
Albrechtsen R, et al. ADAM12 redistributes and activates MMP-14, resulting in gelatin degradation, reduced apoptosis and increased tumor growth. J Cell Sci. 2013;126(Pt 20):4707–20.PubMedCrossRef
75.
Li Y, et al. The overexpression membrane type 1 matrix metalloproteinase is associated with the progression and prognosis in breast cancer. Am J Transl Res. 2015;7(1):120–7.PubMedPubMedCentral
76.
Lodillinsky C, et al. p63/MT1-MMP axis is required for in situ to invasive transition in basal-like breast cancer. Oncogene. 2016;35(3):344–57.PubMedCrossRef
77.
78.
Metadata
Title
Less is more: low expression of MT1-MMP is optimal to promote migration and tumourigenesis of breast cancer cells
Authors
Mario A. Cepeda
Jacob J. H. Pelling
Caitlin L. Evered
Karla C. Williams
Zoey Freedman
Ioana Stan
Jessica A. Willson
Hon S. Leong
Sashko Damjanovski
Publication date
01-12-2016
Publisher
BioMed Central
Published in
Molecular Cancer / Issue 1/2016
Electronic ISSN: 1476-4598
DOI
https://doi.org/10.1186/s12943-016-0547-x

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