Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Article
  • Published:

p140Cap suppresses the invasive properties of highly metastatic MTLn3-EGFR cells via impaired cortactin phosphorylation

Oncogene volume 31, pages 624–633 (2012)Cite this article

Abstract

We have recently shown that the adaptor protein p140Cap regulates tumor properties in terms of cell motility and growth. Here, by using the highly metastatic rat adenocarcinoma cell line MTLn3-epidermal growth factor receptor (EGFR), we assess the role of p140Cap in metastasis formation. Orthotopic transplantation of MTLn3-EGFR cells over-expressing p140Cap in Rag2−/−γc−/− mice resulted in normal primary tumor growth compared with the controls. Strikingly, p140Cap over-expression causes an 80% inhibition in the number of lung metastases. p140Cap over-expressing cells display a 50% reduction in directional cell migration, an increased number and size of focal adhesions, and a strong impairment in the ability to invade in a 3D matrix. p140Cap over-expression affects EGFR signaling and tyrosine phosphorylation of cortactin in response to EGF stimulation. Intriguingly, p140Cap associates with cortactin via interaction with its second proline-rich domain to the cortactin SH3 domain. The phosphomimetic cortactin tyrosine 421 mutant rescues migration and invasive properties in p140Cap over-expressing cells. Taken together, these data demonstrate that p140Cap suppresses the invasive properties of highly metastatic breast carcinoma cells by inhibiting cortactin-dependent cell motility.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  • Ayala I, Baldassarre M, Giacchetti G, Caldieri G, Tete S, Luini A et al. (2008). Multiple regulatory inputs converge on cortactin to control invadopodia biogenesis and extracellular matrix degradation. J Cell Sci 121: 369–378.

    Article  CAS  PubMed  Google Scholar 

  • Baldassarre M, Ayala I, Beznoussenko G, Giacchetti G, Machesky LM, Luini A et al. (2006). Actin dynamics at sites of extracellular matrix degradation. Eur J Cell Biol 85: 1217–1231.

    Article  CAS  PubMed  Google Scholar 

  • Bowden ET, Coopman PJ, Mueller SC . (2001). Invadopodia: unique methods for measurement of extracellular matrix degradation in vitro. Methods Cell Biol 63: 613–627.

    Article  CAS  PubMed  Google Scholar 

  • Buccione R, Caldieri G, Ayala I . (2009). Invadopodia: specialized tumour cell structures for the focal degradation of the extracellular matrix. Cancer Metastasis Rev 28: 137–149.

    Article  PubMed  Google Scholar 

  • Cabodi S, Camacho Leal MP, Di Stefano P, Defilippi P . (2010). Integrin signalling adaptors: not only figurants in the cancer story. Nat Rev Cancer 2010: 858–870.

    Article  Google Scholar 

  • Chin LS, Nugent RD, Raynor MC, Vavalle JP, Li L . (2000). SNIP, a novel SNAP-25-interacting protein implicated in regulated exocytosis. J Biol Chem 275: 1191–1200.

    Article  CAS  PubMed  Google Scholar 

  • Citri A, Yarden Y . (2006). EGF-ERBB signalling: towards the systems level. Nat Rev Mol Cell Biol 7: 505–516.

    Article  CAS  PubMed  Google Scholar 

  • Condeelis J, Segall JE . (2003). Intravital imaging of cell movement in tumours. Nat Rev Cancer 3: 921–930.

    Article  CAS  PubMed  Google Scholar 

  • Damiano L, Di Stefano P, Camacho Leal MP, Barba M, Mainiero F, Cabodi S et al. (2010). p140Cap dual regulation of E-cadherin/EGFR cross-talk and Ras signalling in tumour cell scatter and proliferation. Oncogene 29: 3677–3690.

    Article  CAS  PubMed  Google Scholar 

  • Di Stefano P, Cabodi S, Boeri Erba E, Margaria V, Bergatto E, Giuffrida MG et al. (2004). P130Cas-associated protein (p140Cap) as a new tyrosine-phosphorylated protein involved in cell spreading. Mol Biol Cell 15: 787–800.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Di Stefano P, Damiano L, Cabodi S, Aramu S, Tordella L, Praduroux A et al. (2007). p140Cap protein suppresses tumour cell properties, regulating Csk and Src kinase activity. EMBO J 26: 2843–2855.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gardel ML, Schneider IC, Aratyn-Schaus Y, Waterman CM . (2010). Mechanical integration of actin and adhesion dynamics in cell migration. Annu Rev Cell Dev Biol 26: 315–333.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Head JA, Jiang D, Li M, Zorn LJ, Schaefer EM, Parsons JT et al. (2003). Cortactin tyrosine phosphorylation requires Rac1 activity and association with the cortical actin cytoskeleton. Mol Biol Cell 14: 3216–3229.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ilic D, Furuta Y, Kanazawa S, Takeda N, Sobue K, Nakatsuji N et al. (1995). Reduced cell motility and enhanced focal adhesion contact formation in cells from FAK-deficient mice. Nature 377: 539–544.

    Article  CAS  PubMed  Google Scholar 

  • Jaworski J, Kapitein LC, Gouveia SM, Dortland BR, Wulf PS, Grigoriev I et al. (2009). Dynamic microtubules regulate dendritic spine morphology and synaptic plasticity. Neuron 61: 85–100.

    Article  CAS  PubMed  Google Scholar 

  • Le Devedec SE, van Roosmalen W, Maria N, Grimbergen M, Pont C, Lalai R et al. (2009). An improved model to study tumour cell autonomous metastasis programs using MTLn3 cells and the Rag2(−/−) gammac (−/−) mouse. Clin Exp Metastasis 26: 673–684.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li Y, Tondravi M, Liu J, Smith E, Haudenschild CC, Kaczmarek M et al. (2001). Cortactin potentiates bone metastasis of breast cancer cells. Cancer Res 61: 6906–6911.

    CAS  PubMed  Google Scholar 

  • Lim ST, Mikolon D, Stupack DG, Schlaepfer DD . (2008). FERM control of FAK function: implications for cancer therapy. Cell Cycle 7: 2306–2314.

    Article  CAS  PubMed  Google Scholar 

  • Lorenz M, DesMarais V, Macaluso F, Singer RH, Condeelis J . (2004). Measurement of barbed ends, actin polymerization, and motility in live carcinoma cells after growth factor stimulation. Cell Motil Cytoskeleton 57: 207–217.

    Article  CAS  PubMed  Google Scholar 

  • Lua BL, Low BC . (2005). Cortactin phosphorylation as a switch for actin cytoskeletal network and cell dynamics control. FEBS Lett 579: 577–585.

    Article  CAS  PubMed  Google Scholar 

  • Mader CC, Oser M, Magalhaes MA, Bravo-Cordero JJ, Condeelis J, Koleske AJ, Gil-Henn H . (2011). An EGFR-Src-Arg-cortactin pathway mediates functional maturation of invadopodia and breast cancer cell invasion. Cancer Res 71: 1730–1741.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Marxfeld H, Staedtler F, Harleman JH . (2006). Characterisation of two rat mammary tumour models for breast cancer research by gene expression profiling. Exp Toxicol Pathol 58: 133–143.

    Article  CAS  PubMed  Google Scholar 

  • Mori S, Chang JT, Andrechek ER, Matsumura N, Baba T, Yao G et al. (2009). Anchorage-independent cell growth signature identifies tumours with metastatic potential. Oncogene 28: 2796–2805.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Neri A, Nicolson GL . (1981). Phenotypic drift of metastatic and cell-surface properties of mammary adenocarcinoma cell clones during growth in vitro. Int J Cancer 28: 731–738.

    Article  CAS  PubMed  Google Scholar 

  • Normanno N, De Luca A, Bianco C, Strizzi L, Mancino M, Maiello MR et al. (2006). Epidermal growth factor receptor (EGFR) signalling in cancer. Gene 366: 2–16.

    Article  CAS  PubMed  Google Scholar 

  • Oser M, Yamaguchi H, Mader CC, Bravo-Cordero JJ, Arias M, Chen X et al. (2009). Cortactin regulates cofilin and N-WASp activities to control the stages of invadopodium assembly and maturation. J Cell Biol 186: 571–587.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sahai E . (2007). Illuminating the metastatic process. Nat Rev Cancer 7: 737–749.

    Article  CAS  PubMed  Google Scholar 

  • Vicente-Manzanares M, Webb DJ, Horwitz AR . (2005). Cell migration at a glance. J Cell Sci 118: 4917–4919.

    Article  CAS  PubMed  Google Scholar 

  • Weaver AM . (2008). Cortactin in tumour invasiveness. Cancer Lett 265: 157–166.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Webb DJ, Zhang H, Horwitz AF . (2005). Cell migration: an overview. Methods Mol Biol 294: 3–11.

    PubMed  Google Scholar 

  • Weed SA, Karginov AV, Schafer DA, Weaver AM, Kinley AW, Cooper JA et al. (2000). Cortactin localization to sites of actin assembly in lamellipodia requires interactions with F-actin and the Arp2/3 complex. J Cell Biol 151: 29–40.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wolfenson H, Henis YI, Geiger B, Bershadsky AD . (2009). The heel and toe of the cell's foot: a multifaceted approach for understanding the structure and dynamics of focal adhesions. Cell Motil Cytoskeleton 66: 1017–1029.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wu H, Parsons JT . (1993). Cortactin, an 80/85-kilodalton pp60src substrate, is a filamentous actin-binding protein enriched in the cell cortex. J Cell Biol 120: 1417–1426.

    Article  CAS  PubMed  Google Scholar 

  • Wu H, Reynolds AB, Kanner SB, Vines RR, Parsons JT . (1991). Identification and characterization of a novel cytoskeleton-associated pp60src substrate. Mol Cell Biol 11: 5113–5124.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xue C, Wyckoff J, Liang F, Sidani M, Violini S, Tsai KL et al. (2006). Epidermal growth factor receptor overexpression results in increased tumour cell motility in vivo coordinately with enhanced intravasation and metastasis. Cancer Res 66: 192–197.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank Stefan Huveneers for help with the in vitro angiogenesis assay. This work was supported by EU FP7 Metafight project, AIRC, MUR (PRIN), Regione Piemonte – Progetti Sanità, Oncoprot, PiSTEM, Druidi and CIPE, Compagnia San Paolo, Torino, the Dutch Cancer Society (UL 2006-3521, UL 2007-3860 and UL 2010-4670) and the Netherlands Genomics Initiative grant to the Netherlands Toxicogenomics Center grant.

Author information

Author notes

  1. L Damiano, S E Le Dévédec and P Di Stefano: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Genetics, Molecular Biotechnology Centre, Biology and Biochemistry, University of Torino, Torino, Italy

    L Damiano, P Di Stefano, D Repetto, E De Luca, E Turco & P Defilippi

  2. Division of Toxicology, Leiden Amsterdam Center for Drug Research (LACDR), Leiden University, Leiden, The Netherlands

    S E Le Dévédec, R Lalai, H Truong, J L Xiong, E H Danen & B van de Water

  3. Imaging and BioInformatics, Leiden Institute of Advanced Computer Science, Leiden University, Leiden, The Netherlands

    K Yan & F J Verbeek

  4. Tumour Cell Invasion Laboratory, Consorzio Mario Negri Sud, Chieti, Italy

    F Attanasio & R Buccione

Authors
  1. L Damiano
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S E Le Dévédec
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. P Di Stefano
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D Repetto
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. R Lalai
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. H Truong
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. J L Xiong
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. E H Danen
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. K Yan
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. F J Verbeek
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. E De Luca
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. F Attanasio
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. R Buccione
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. E Turco
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. B van de Water
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. P Defilippi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to P Defilippi.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies the paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Damiano, L., Le Dévédec, S., Di Stefano, P. et al. p140Cap suppresses the invasive properties of highly metastatic MTLn3-EGFR cells via impaired cortactin phosphorylation. Oncogene 31, 624–633 (2012). https://doi.org/10.1038/onc.2011.257

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2011.257

Keywords

This article is cited by

Search

Advanced search

Quick links