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Upregulation of Eps8 in oral squamous cell carcinoma promotes cell migration and invasion through integrin-dependent Rac1 activation

Oncogene volume 28, pages 2524–2534 (2009)Cite this article

Abstract

Oral squamous cell carcinoma (OSCC) is a lethal disease and early death usually occurs as a result of local invasion and regional lymph node metastases. Current treatment regimens are, to a certain degree, inadequate, with a 5-year mortality rate of around 50% and novel therapeutic targets are urgently required. Using expression microarrays, we identified the eps8 gene as being overexpressed in OSCC cell lines relative to normal oral keratinocytes, and confirmed these findings using RT–PCR and western blotting. In human tissues, we found that Eps8 was upregulated in OSCC (32% of primary tumors) compared with normal oral mucosa, and that expression correlated significantly with lymph node metastasis (P=0.032), suggesting a disease-promoting effect. Using OSCC cell lines, we assessed the functional role of Eps8 in tumor cells. Although suppression of Eps8 produced no effect on cell proliferation, both cell spreading and migration were markedly inhibited. The latter cell functions may be modulated through the small GTP-ase, Rac1 and we used pull-down assays to investigate the role of Eps8 in Rac1 signaling. We found that αvβ6- and α5β1-integrin-dependent activation of Rac1 was mediated through Eps8. Knockdown of either Eps8 or Rac1, inhibited integrin-dependent cell migration similarly and transient expression of constitutively active Rac1 restored migration of cells in which Eps8 expression had been suppressed. We also showed that knockdown of Eps8 inhibited tumor cell invasion in an organotypic model of OSCC. These data suggest that Eps8 and Rac1 are part of an integrated signaling pathway modulating integrin-dependent tumour cell motility and identify Eps8 as a possible therapeutic target.

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References

  • Alevizos I, Mahadevappa M, Zhang X, Ohyama H, Kohno Y, Posner M et al. (2001). Oral cancer in vivo gene expression profiling assisted by laser capture microdissection and microarray analysis. Oncogene 20: 6196–6204.

    Article  CAS  Google Scholar 

  • Al Moustafa AE, Alaoui-Jamali MA, Batist G, Hernandez-Perez M, Serruya C, Alpert L et al. (2002). Identification of genes associated with head and neck carcinogenesis by cDNA microarray comparison between matched primary normal epithelial and squamous carcinoma cells. Oncogene 21: 2634–2640.

    Article  Google Scholar 

  • Bates RC, Bellovin DI, Brown C, Maynard E, Wu B, Kawakatsu H et al. (2005). Transcriptional activation of integrin β6 during the epithelial-mesenchymal transition defines a novel prognostic indicator of aggressive colon carcinoma. J Clin Invest 115: 339–347.

    Article  CAS  Google Scholar 

  • Croce A, Cassata G, Disanza A, Gagliani MC, Tacchetti C, Malabarba MG et al. (2004). A novel actin barbed-end-capping activity in EPS-8 regulates apical morphogenesis in intestinal cells of Caenorhabditis elegans. Nat Cell Biol 6: 1173–1179.

    Article  CAS  Google Scholar 

  • Cromer A, Carles A, Millon R, Ganguli G, Chalmel F, Lemaire F et al. (2004). Identification of genes associated with tumorigenesis and metastatic potential of hypopharyngeal cancer by microarray analysis. Oncogene 23: 2484–2498.

    Article  CAS  Google Scholar 

  • del Pozo MA, Price LS, Alderson NB, Ren XD, Schwartz MA . (2000). Adhesion to the extracellular matrix regulates the coupling of the small GTPase Rac to its effector PAK. EMBO J 19: 2008–2014.

    Article  CAS  Google Scholar 

  • del Pozo MA, Alderson NB, Kiosses WB, Chiang HH, Anderson RG, Schwartz MA . (2004). Integrins regulate Rac targeting by internalization of membrane domains. Science 303: 839–842.

    Article  CAS  Google Scholar 

  • Disanza A, Carlier MF, Stradal TE, Didry D, Frittoli E, Confalonieri S et al. (2004). Eps8 controls actin-based motility by capping the barbed ends of actin filaments. Nat Cell Biol 6: 1180–1188.

    Article  CAS  Google Scholar 

  • Dormond O, Foletti A, Paroz C, Ruegg C . (2001). NSAIDS inhibit αvβ3 integrin-mediated and Cdc/Rac-dependent endothelial-cell spreading, migration and angiogenesis. Nat Med 7: 1041–1047.

    Article  CAS  Google Scholar 

  • Elayadi AN, Samli KN, Prudkin L, Liu YH, Bian A, Xie XJ et al. (2007). A peptide selected by biopanning identifies the integrin αvβ6 as a prognostic biomarker for non-small cell lung cancer. Cancer Res 67: 5889–5895.

    Article  CAS  Google Scholar 

  • Fazioli F, Minichiello L, Matoska V, Castagnino P, Miki T, Wong WT et al. (1993). Eps8, a substrate for the epidermal growth factor receptor kinase, enhances EGF-dependent mitogenic signals. EMBO J 12: 3799–3808.

    Article  CAS  Google Scholar 

  • Funato Y, Terabayashi T, Suenaga N, Seiki M, Takenawa T, Miki H . (2004). IRSp53/Eps8 complex is important for positive regulation of Rac and cancer cell motility/invasiveness. Cancer Res 64: 5237–5244.

    Article  CAS  Google Scholar 

  • Innocenti M, Frittoli E, Ponzanelli I, Falck JR, Brachmann SM, Di Fiore PP et al. (2003). Phosphoinositide 3-kinase activates Rac by entering in a complex with Eps8, Abi1, and Sos-1. J Cell Biol 160: 17–23.

    Article  CAS  Google Scholar 

  • Inobe M, Katsube K, Miyagoe Y, Nabeshima Y, Takeda S . (1999). Identification of EPS8 as a Dvl1-associated molecule. Biochem Biophys Res Commun 266: 216–221.

    Article  CAS  Google Scholar 

  • Irizarry RA, Bolstad BM, Collin F, Cope LM, Hobbs B, Speed TP . (2003). Summaries of Affymetrix GeneChip probe level data. Nucleic Acids Res 31: e15.

    Article  Google Scholar 

  • Jeon GA, Lee JS, Patel V, Gutkind JS, Thorgeirsson SS, Kim EC et al. (2004). Global gene expression profiles of human head and neck squamous carcinoma cell lines. Int J Cancer 112: 249–258.

    Article  CAS  Google Scholar 

  • Kim M, Califano J . (2004). Molecular Pathology of Head and Neck Cancer. Int J Cancer 112: 545–553.

    Article  CAS  Google Scholar 

  • Maa MC, Hsieh CY, Leu TH . (2001). Overexpression of p97Eps8 leads to cellular transformation: implication of pleckstrin homology domain in p97Eps8-mediated ERK activation. Oncogene 20: 106–112.

    Article  CAS  Google Scholar 

  • Maa MC, Lee JC, Chen YJ, Chen YJ, Lee YC, Wang ST et al. (2007). EPS8 facilitates cellular growth and motility of colon cancer cells by increasing the expression and activity of focal adhesion kinase. J Biol Chem 282: 19399–19409.

    Article  CAS  Google Scholar 

  • Marsh D, Dickinson S, Neill GW, Marshall JF, Hart IR, Thomas GJ . (2008). alphavbeta6 integrin promotes invasion of morphoeic basal cell carcinoma through stromal modulation. Cancer Res 68: 3295–3303.

    Article  CAS  Google Scholar 

  • Matoskova B, Wong WT, Salcini AE, Pelicci PG, Di Fiore PP . (1995). Constitutive phosphorylation of Eps8 in tumor cell lines: relevance to malignant transformation. Mol Cell Biol 15: 3805–3812.

    Article  CAS  Google Scholar 

  • Miki H, Yamaguchi H, Suetsugu S, Takenawa T . (2000). IRSp53 is an essential intermediate between Rac and WAVE in the regulation of membrane ruffling. Nature 408: 732–735.

    Article  CAS  Google Scholar 

  • Myers JN, Greenberg JS, Mo V, Roberts D . (2001). Extracapsular spread. A significant predictor of treatment failure in patients with squamous cell carcinoma of the tongue. Cancer 92: 3030–3036.

    Article  CAS  Google Scholar 

  • Nystrom ML, Thomas GJ, Hart IR, Stone M, McKenzie IC, Marshall JF . (2005). Development of a quantitative method to analyse tumor cell invasion in organotypic culture. J Pathol 205: 468–475.

    Article  CAS  Google Scholar 

  • Nystrom ML, McCulloch D, Weinreb PH, Violette SM, Speight PM, Marshall JF et al. (2006). Cyclooxygenase-2 inhibition suppresses alphavbeta6 integrin-dependent oral squamous carcinoma invasion. Cancer Res 66: 10833–41082.

    Article  CAS  Google Scholar 

  • Parkin DM, Bray F, Ferlay J, Pisani P . (2005). Global cancer statistics 2002. CA Cancer J Clin 55: 74–108.

    Article  Google Scholar 

  • Prime SS, Nixon SV, Crane IJ, Stone A, Matthews JB, Maitland NJ et al. (1990). The behaviour of human oral squamous cell carcinoma in cell culture. J Pathol 160: 259–269.

    Article  CAS  Google Scholar 

  • Scita G, Nordstrom J, Carbone R, Tenca P, Giardina G, Gutkind S et al. (1999). EPS8 and E3B1 transduce signals from Ras to Rac. Nature 401: 290–293.

    Article  CAS  Google Scholar 

  • Scita G, Tenca P, Areces LB, Tocchetti A, Frittoli E, Giardina G et al. (2001). An effector region in Eps8 is responsible for the activation of the Rac-specific GEF activity of Sos-1 and for the proper localization of the Rac-based actin-polymerizing machine. J Cell Biol 154: 1031–1044.

    Article  CAS  Google Scholar 

  • Ramsay AG, Keppler MD, Jazayeri M, Thomas GJ, Parsons M, Violette S et al. (2007). HS1-associated protein X-1 regulates carcinoma cell migration and invasion via clathrin-mediated endocytosis of integrin alphavbeta6. Cancer Res 67: 5275–5284.

    Article  CAS  Google Scholar 

  • Thirthagiri E, Robinson CM, Huntley S, Davies M, Yap LF, Prime SS et al. (2007). Spindle assembly checkpoint and centrosome abnormalities in oral cancer. Cancer Lett 258: 276–285.

    Article  CAS  Google Scholar 

  • Thomas GJ, Hart IR, Speight PM, Marshall JF . (2002). Binding of TGF-beta1 latency-associated peptide (LAP) to alpha(v)beta6 integrin modulates behaviour of squamous carcinoma cells. Br J Cancer 87: 859–867.

    Article  CAS  Google Scholar 

  • Thomas GJ, Lewis MP, Hart IR, Marshall JF, Speight PM . (2001). AlphaVbeta6 integrin promotes invasion of squamous carcinoma cells through up-regulation of matrix metalloproteinase-9. Int J Cancer 92: 641–650.

    Article  CAS  Google Scholar 

  • Tusher VG, Tibshirani R, Chu G . (2001). Significance analysis of microarrays applied to the ionizing radiation response. Proc Natl Acad Sci USA 98: 5116–5121.

    Article  CAS  Google Scholar 

  • Welsch T, Endlich K, Giese T, Buchler MW, Schmidt J . (2007). Eps8 is increased in pancreatic cancer and required for dynamic actin-based cell protrusions and intercellular cytoskeletal organization. Cancer Lett 255: 205–218.

    Article  CAS  Google Scholar 

  • Woolgar JA, Rogers SN, Lowe D, Brown JS, Vaughan ED . (2003). Cervical lymph node metastasis in oral cancer: the importance of even microscopic extracapsular spread. Oral Oncol 39: 130–137.

    Article  CAS  Google Scholar 

  • Yao J, Weremowicz S, Feng B, Gentleman RC, Marks JR, Gelman R et al. (2006). Combined cDNA array comparative genomic hybridization and serial analysis of gene expression analysis of breast tumor progression. Cancer Res 66: 4065–4078.

    Article  CAS  Google Scholar 

  • Yeudall WA, Miyazaki H, Ensley JF, Cardinali M, Gutkind JS, Patel V . (2005). Uncoupling of epidermal growth factor-dependent proliferation and invasion in a model of squamous carcinoma progression. Oral Oncol 41: 698–708.

    Article  CAS  Google Scholar 

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Acknowledgements

The Cancer Research Initiatives Foundation, Malaysia and The Health Foundation supported the research.

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Author notes

  1. L F Yap and V Jenei: These authors contributed equally to this work.

  2. G J Thomas and I C Paterson: These authors jointly supervised this study.

Authors and Affiliations

  1. Department of Oral and Dental Science, University of Bristol, Bristol, UK

    L F Yap, T M Benn, S P Threadgold & I C Paterson

  2. Cancer Research Initiatives Foundation, Kuala Lumpur, Malaysia

    L F Yap

  3. Centre for Tumor Biology, Institute of Cancer, Bart's and the London School of Medicine and Dentistry, London, UK

    V Jenei, K Moutasim & G J Thomas

  4. Cancer Sciences Division, Southampton University School of Medicine, Southampton, UK

    V Jenei, K Moutasim & G J Thomas

  5. School of Dental Sciences, University of Newcastle, Newcastle, UK

    C M Robinson

  6. Edinburgh Dental Institute, University of Edinburgh, Edinburgh, UK

    V Lopes

  7. Institute for Cancer Studies, University of Birmingham, Birmingham, UK

    W Wei

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  1. L F Yap
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Correspondence to G J Thomas.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)

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Yap, L., Jenei, V., Robinson, C. et al. Upregulation of Eps8 in oral squamous cell carcinoma promotes cell migration and invasion through integrin-dependent Rac1 activation. Oncogene 28, 2524–2534 (2009). https://doi.org/10.1038/onc.2009.105

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