Membrane Type 1 Matrix Metalloproteinase induces an epithelial to mesenchymal transition and cancer stem cell-like properties in SCC9 cells

BMC Cancer

Cong-Chong Yang²  Li-Fang Zhu⁴  Xiao-Hui Xu¹  Tian-Yun Ning²  Jin-Hai Ye³  Lai-Kui Liu² 

[1] Department of the First Outpatient, College of Stomatology, Nanjing Medical University, Nanjing, People’s Republic of China

[2] Department of Basic Science of Stomatology, College of Stomatology, Nanjing Medical University, Postal# 210029 136# Hanzhong Road, Nanjing, Jiangsu, People’s Republic of China

[3] Department of Oral and Maxillofacial Surgery, College of Stomatology, Nanjing Medical University, Nanjing, People’s Republic of China

[4] Department of Stomatology, the First Affiliated Hospital of Soochow University, Suzhou, People’s Republic of China

关键词: Oral squamous cell carcinoma;    Cancer stem cell;    EMT;    Membrane type 1 matrix metalloproteinase;   

Others  :  1079819
DOI  :  10.1186/1471-2407-13-171

 received in 2012-08-19, accepted in 2013-01-30,  发布年份 2013

【 摘 要 】

Background

Tissue invasion and metastasis are acquired abilities of cancer and related to the death in oral squamous cell carcinoma (OSCC). Emerging observations indicate that the epithelial-to-mesenchymal transition (EMT) is associated with tumor progression and the generation of cells with cancer stem cells (CSCs) properties. Membrane Type 1 Matrix Metalloproteinase (MT1-MMP) is a cell surface proteinase, which is involved in degrading extracellular matrix components that can promote tumor invasion and cell migration.

Methods

In the current study, we utilized SCC9 cells stably transfected with an empty vector (SCC9-N) or a vector encoding human MT1-MMP (SCC9-M) to study the role of MT1-MMP in EMT development.

Results

Upon up-regulation of MT1-MMP, SCC9-M cells underwent EMT, in which they presented a fibroblast-like phenotype and had a decreased expression of epithelial markers (E-cadherin, cytokeratin18 and β-catenin) and an increased expression of mesenchymal markers (vimentin and fibronectin). We further demonstrated that MT1-MMP-induced morphologic changes increased the level of Twist and ZEB, and were dependent on repressing the transcription of E-cadherin. These activities resulted in low adhesive, high invasive abilities of the SCC9-M cells. Furthermore, MT1-MMP-induced transformed cells exhibited cancer stem cell (CSC)-like characteristics, such as low proliferation, self-renewal ability, resistance to chemotherapeutic drugs and apoptosis, and expression of CSCs surface markers.

Conclusions

In conclusion, our study indicates that overexpression of MT1-MMP induces EMT and results in the acquisition of CSC-like properties in SCC9 cells. Our growing understanding of the mechanism regulating EMT may provide new targets against invasion and metastasis in OSCC.

【 授权许可】

   
2013 Yang et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Forastiere A, Koch W, Trotti A, Sidransky D: Head and neck cancer. N Engl J Med 2001, 345(26):1890-1900.
• [2]Carvalho AL, Nishimoto IN, Califano JA, Kowalski LP: Trends in incidence and prognosis for head and neck cancer in the United States: a site-specific analysis of the SEER database. International journal of cancer Journal international du cancer 2005, 114(5):806-816.
• [3]Hanahan D, Weinberg RA: The hallmarks of cancer. Cell 2000, 100(1):57-70.
• [4]Prince ME, Sivanandan R, Kaczorowski A, Wolf GT, Kaplan MJ, Dalerba P, Weissman IL, Clarke MF, Ailles LE: Identification of a subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma. Proc Natl Acad Sci USA 2007, 104(3):973-978.
• [5]Thiery JP: Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer 2002, 2(6):442-454.
• [6]Huber MA, Kraut N, Beug H: Molecular requirements for epithelial-mesenchymal transition during tumor progression. Curr Opin Cell Biol 2005, 17(5):548-558.
• [7]Kalluri R, Neilson EG: Epithelial-mesenchymal transition and its implications for fibrosis. J Clin Invest 2003, 112(12):1776-1784.
• [8]Kalluri R, Weinberg RA: The basics of epithelial-mesenchymal transition. J Clin Invest 2009, 119(6):1420-1428.
• [9]Thiery JP, Acloque H, Huang RY, Nieto MA: Epithelial-mesenchymal transitions in development and disease. Cell 2009, 139(5):871-890.
• [10]Nieto MA: The ins and outs of the epithelial to mesenchymal transition in health and disease. Annu Rev Cell Dev Biol 2011, 27:347-376.
• [11]Peinado H, Olmeda D, Cano A: Snail, Zeb and bHLH factors in tumour progression: an alliance against the epithelial phenotype? Nat Rev Cancer 2007, 7(6):415-428.
• [12]Brinckerhoff CE, Matrisian LM: Matrix metalloproteinases: a tail of a frog that became a prince. Nat Rev Mol Cell Biol 2002, 3(3):207-214.
• [13]Page-McCaw A, Ewald AJ, Werb Z: Matrix metalloproteinases and the regulation of tissue remodelling. Nat Rev Mol Cell Biol 2007, 8(3):221-233.
• [14]Seiki M: Membrane-type matrix metalloproteinases. APMIS: acta pathologica, microbiologica, et immunologica Scandinavica 1999, 107(1):137-143.
• [15]Itoh Y, Nagase H: Matrix metalloproteinases in cancer. Essays Biochem 2002, 38:21-36.
• [16]Sato H, Takino T, Miyamori H: Roles of membrane-type matrix metalloproteinase-1 in tumor invasion and metastasis. Cancer Sci 2005, 96(4):212-217.
• [17]Itoh Y: MT1-MMP: a key regulator of cell migration in tissue. IUBMB Life 2006, 58(10):589-596.
• [18]Seiki M: The cell surface: the stage for matrix metalloproteinase regulation of migration. Curr Opin Cell Biol 2002, 14(5):624-632.
• [19]Zarrabi K, Dufour A, Li J, Kuscu C, Pulkoski-Gross A, Zhi J, Hu Y, Sampson NS, Zucker S, Cao J: Inhibition of matrix metalloproteinase 14 (MMP-14)-mediated cancer cell migration. J Biol Chem 2011, 286(38):33167-33177.
• [20]Perentes JY, Kirkpatrick ND, Nagano S, Smith EY, Shaver CM, Sgroi D, Garkavtsev I, Munn LL, Jain RK, Boucher Y: Cancer cell-associated MT1-MMP promotes blood vessel invasion and distant metastasis in triple-negative mammary tumors. Cancer Res 2011, 71(13):4527-4538.
• [21]Rozanov DV, Deryugina EI, Monosov EZ, Marchenko ND, Strongin AY: Aberrant, persistent inclusion into lipid rafts limits the tumorigenic function of membrane type-1 matrix metalloproteinase in malignant cells. Exp Cell Res 2004, 293(1):81-95.
• [22]Cao J, Chiarelli C, Richman O, Zarrabi K, Kozarekar P, Zucker S: Membrane type 1 matrix metalloproteinase induces epithelial-to-mesenchymal transition in prostate cancer. J Biol Chem 2008, 283(10):6232-6240.
• [23]Reya T, Morrison SJ, Clarke MF, Weissman IL: Stem cells, cancer, and cancer stem cells. Nature 2001, 414(6859):105-111.
• [24]Spaderna S, Schmalhofer O, Hlubek F, Berx G, Eger A, Merkel S, Jung A, Kirchner T, Brabletz T: A transient, EMT-linked loss of basement membranes indicates metastasis and poor survival in colorectal cancer. Gastroenterology 2006, 131(3):830-840.
• [25]Mani SA, Guo W, Liao MJ, Eaton EN, Ayyanan A, Zhou AY, Brooks M, Reinhard F, Zhang CC, Shipitsin M: The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell 2008, 133(4):704-715.
• [26]Waldmann J, Slater EP, Langer P, Buchholz M, Ramaswamy A, Walz MK, Schmid KW, Feldmann G, Bartsch DK, Fendrich V: Expression of the transcription factor snail and its target gene twist are associated with malignancy in pheochromocytomas. Ann Surg Oncol 2009, 16(7):1997-2005.
• [27]Mosmann T: Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983, 65(1–2):55-63.
• [28]Foroni C, Broggini M, Generali D, Damia G: Epithelial-mesenchymal transition and breast cancer: Role, molecular mechanisms and clinical impact. Cancer Treat Rev 2012, 38(6):689-697.
• [29]Kam Y, Guess C, Estrada L, Weidow B, Quaranta V: A novel circular invasion assay mimics in vivo invasive behavior of cancer cell lines and distinguishes single-cell motility in vitro. BMC Cancer 2008, 8:198. BioMed Central Full Text
• [30]Li J, Zhou BP: Activation of beta-catenin and Akt pathways by Twist are critical for the maintenance of EMT associated cancer stem cell-like characters. BMC Cancer 2011, 11:49. BioMed Central Full Text
• [31]Dean M, Fojo T, Bates S: Tumour stem cells and drug resistance. Nat Rev Cancer 2005, 5(4):275-284.
• [32]Chen C, Wei Y, Hummel M, Hoffmann TK, Gross M, Kaufmann AM, Albers AE: Evidence for epithelial-mesenchymal transition in cancer stem cells of head and neck squamous cell carcinoma. PLoS One 2011, 6(1):e16466.
• [33]Krantz SB, Shields MA, Dangi-Garimella S, Munshi HG, Bentrem DJ: Contribution of epithelial-to-mesenchymal transition and cancer stem cells to pancreatic cancer progression. J Surg Res 2012, 173(1):105-112.
• [34]Hotary K, Li XY, Allen E, Stevens SL, Weiss SJ: A cancer cell metalloprotease triad regulates the basement membrane transmigration program. Genes Dev 2006, 20(19):2673-2686.
• [35]Ries C, Egea V, Karow M, Kolb H, Jochum M, Neth P: MMP-2, MT1-MMP, and TIMP-2 are essential for the invasive capacity of human mesenchymal stem cells: differential regulation by inflammatory cytokines. Blood 2007, 109(9):4055-4063.
• [36]Peinado H, Cano A: New potential therapeutic targets to combat epithelial tumor invasion. Clinical & translational oncology: official publication of the Federation of Spanish Oncology Societies and of the National Cancer Institute of Mexico 2006, 8(12):851-857.
• [37]Cano A, Perez-Moreno MA, Rodrigo I, Locascio A, Blanco MJ, del Barrio MG, Portillo F, Nieto MA: The transcription factor snail controls epithelial-mesenchymal transitions by repressing E-cadherin expression. Nat Cell Biol 2000, 2(2):76-83.
• [38]Higashikawa K, Yoneda S, Taki M, Shigeishi H, Ono S, Tobiume K, Kamata N: Gene expression profiling to identify genes associated with high-invasiveness in human squamous cell carcinoma with epithelial-to-mesenchymal transition. Cancer Lett 2008, 264(2):256-264.
• [39]Zhu LF, Hu Y, Yang CC, Xu XH, Ning TY, Wang ZL, Ye JH, Liu LK: Snail overexpression induces an epithelial to mesenchymal transition and cancer stem cell-like properties in SCC9 cells. Laboratory investigation; a journal of technical methods and pathology 2012, 92(5):744-752.
• [40]Yang J, Mani SA, Donaher JL, Ramaswamy S, Itzykson RA, Come C, Savagner P, Gitelman I, Richardson A, Weinberg RA: Twist, a master regulator of morphogenesis, plays an essential role in tumor metastasis. Cell 2004, 117(7):927-939.
• [41]Yang J, Mani SA, Weinberg RA: Exploring a new twist on tumor metastasis. Cancer Res 2006, 66(9):4549-4552.
• [42]Sanchez-Tillo E, de Barrios O, Siles L, Cuatrecasas M, Castells A, Postigo A: beta-catenin/TCF4 complex induces the epithelial-to-mesenchymal transition (EMT)-activator ZEB1 to regulate tumor invasiveness. Proc Natl Acad Sci USA 2011, 108(48):19204-19209.
• [43]Zhang Z, Filho MS, Nor JE: The biology of head and neck cancer stem cells. Oral Oncol 2012, 48(1):1-9.
• [44]Klarmann GJ, Hurt EM, Mathews LA, Zhang X, Duhagon MA, Mistree T, Thomas SB, Farrar WL: Invasive prostate cancer cells are tumor initiating cells that have a stem cell-like genomic signature. Clin Exp Metastasis 2009, 26(5):433-446.
• [45]Clarke MF, Dick JE, Dirks PB, Eaves CJ, Jamieson CH, Jones DL, Visvader J, Weissman IL, Wahl GM: Cancer stem cells–perspectives on current status and future directions: AACR Workshop on cancer stem cells. Cancer Res 2006, 66(19):9339-9344.