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Characterization of the interface between normal and transformed epithelial cells

Nature Cell Biology volume 11, pages 460–467 (2009)Cite this article

Abstract

In most cancers, transformation begins in a single cell in an epithelial cell sheet1,2,3. However, it is not known what happens at the interface between non-transformed (normal) and transformed cells once the initial transformation has occurred. Using Madin-Darby canine kidney (MDCK) epithelial cells that express constitutively active, oncogenic Ras (RasV12) in a tetracycline-inducible system, we investigated the cellular processes arising at the interface between normal and transformed cells. We show that two independent phenomena occur in a non-cell-autonomous manner: when surrounded by normal cells, RasV12 cells are either apically extruded from the monolayer, or form dynamic basal protrusions and invade the basal matrix. Neither apical extrusion nor basal protrusion formation is observed when RasV12 cells are surrounded by other RasV12 cells. We show that Cdc42 and ROCK (also known as Rho kinase) have vital roles in these processes. We also demonstrate that E-cadherin knockdown in normal cells surrounding RasV12 cells reduces the frequency of apical extrusion, while promoting basal protrusion formation and invasion. These results indicate that RasV12-transformed cells are able to recognize differences between normal and transformed cells, and consequently leave epithelial sheets either apically or basally, in a cell-context-dependent manner.

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Figure 1: Epithelial cells expressing RasV12 are apically extruded from surrounding normal epithelium in a non-cell-autonomous manner.
Figure 2: Molecular mechanism for apical extrusion of RasV12-expressing cells from a monolayer of normal cells.
Figure 3: Non-extruded GFP–RasV12 cells produce dynamic basal protrusions beneath the neighbouring MDCK cells.
Figure 4: Molecular mechanisms for apical extrusion and basal protrusion formation of RasV12 cells in a monolayer of normal cells.
Figure 5: E-cadherin-based intercellular adhesions of surrounding normal cells can influence the fate of RasV12 cells.

References

  1. Hanahan, D. & Weinberg, R. A. The hallmarks of cancer. Cell 100, 57–70 (2000).

    Article  CAS  Google Scholar 

  2. Fialkow, P. J. Clonal origin of human tumors. Biochim. Biophys. Acta. 458, 283–321 (1976).

    CAS  PubMed  Google Scholar 

  3. Nowell, P. C. The clonal evolution of tumor cell populations. Science 194, 23–28 (1976).

    Article  CAS  Google Scholar 

  4. Schoenenberger, C. A., Zuk, A., Kendall, D. & Matlin, K. S. Multilayering and loss of apical polarity in MDCK cells transformed with viral K-ras. J. Cell Biol. 112, 873–889 (1991).

    Article  CAS  Google Scholar 

  5. Rosenblatt, J., Raff, M. C. & Cramer, L. P. An epithelial cell destined for apoptosis signals its neighbors to extrude it by an actin- and myosin-dependent mechanism. Curr. Biol. 11, 1847–1857 (2001).

    Article  CAS  Google Scholar 

  6. Bos, J. L. Ras-like GTPases. Biochim. Biophys. Acta. 1333, M19–31 (1997).

    CAS  PubMed  Google Scholar 

  7. Baena-Lopez, L. A., Pastor-Pareja, J. C. & Resino, J. Wg and Egfr signalling antagonise the development of the peripodial epithelium in Drosophila wing discs. Development 130, 6497–6506 (2003).

    Article  CAS  Google Scholar 

  8. Rodriguez-Viciana, P. et al. Role of phosphoinositide 3-OH kinase in cell transformation and control of the actin cytoskeleton by Ras. Cell 89, 457–467 (1997).

    Article  CAS  Google Scholar 

  9. Cantrell, D. A. Phosphoinositide 3-kinase signalling pathways. J. Cell Sci. 114, 1439–1445 (2001).

    CAS  PubMed  Google Scholar 

  10. Karnoub, A. E. & Weinberg, R. A. Ras oncogenes: split personalities. Nature Rev. Mol. Cell Biol. 9, 517–531 (2008).

    Article  CAS  Google Scholar 

  11. Kimura, K. et al. Regulation of myosin phosphatase by Rho and Rho-associated kinase (Rho-kinase). Science 273, 245–248 (1996).

    Article  CAS  Google Scholar 

  12. Jaffe, A. B. & Hall, A. Rho GTPases: biochemistry and biology. Annu. Rev. Cell Dev. Biol. 21, 247–269 (2005).

    Article  CAS  Google Scholar 

  13. Vega, F. M. & Ridley, A. J. Rho GTPases in cancer cell biology. FEBS Lett. 582, 2093–2101 (2008).

    Article  CAS  Google Scholar 

  14. Sahai, E., Olson, M. F. & Marshall, C. J. Cross-talk between Ras and Rho signalling pathways in transformation favours proliferation and increased motility. EMBO J. 20, 755–766 (2001).

    Article  CAS  Google Scholar 

  15. de la Cova, C., Abril, M., Bellosta, P., Gallant, P. & Johnston, L. A. Drosophila Myc regulates organ size by inducing cell competition. Cell 117, 107–116 (2004).

    Article  CAS  Google Scholar 

  16. Moreno, E. & Basler, K. dMyc transforms cells into super-competitors. Cell 117, 117–129 (2004).

    Article  CAS  Google Scholar 

  17. Vidal, M., Larson, D. E. & Cagan, R. L. Csk-deficient boundary cells are eliminated from normal Drosophila epithelia by exclusion, migration, and apoptosis. Dev. Cell 10, 33–44 (2006).

    Article  CAS  Google Scholar 

  18. Brumby, A. M. & Richardson, H. E. scribble mutants cooperate with oncogenic Ras or Notch to cause neoplastic overgrowth in Drosophila. EMBO J. 22, 5769–5779 (2003).

    Article  CAS  Google Scholar 

  19. Stoker, M. G., Shearer, M. & O'Neill, C. Growth inhibition of polyoma-transformed cells by contact with static normal fibroblasts. J. Cell Sci. 1, 297–310 (1966).

    CAS  PubMed  Google Scholar 

  20. Bignami, M., Rosa, S., La Rocca, S. A., Falcone, G. & Tato, F. Differential influence of adjacent normal cells on the proliferation of mammalian cells transformed by the viral oncogenes myc, ras and src. Oncogene 2, 509–514 (1988).

    CAS  PubMed  Google Scholar 

  21. Alexander, D. B. et al. Normal cells control the growth of neighboring transformed cells independent of gap junctional communication and SRC activity. Cancer Res. 64, 1347–1358 (2004).

    Article  CAS  Google Scholar 

  22. Wells, C. M., Walmsley, M., Ooi, S., Tybulewicz, V. & Ridley, A. J. Rac1-deficient macrophages exhibit defects in cell spreading and membrane ruffling but not migration. J. Cell Sci. 117, 1259–1268 (2004).

    Article  CAS  Google Scholar 

  23. Hogan, C. et al. Rap1 regulates the formation of E-cadherin-based cell-cell contacts. Mol. Cell. Biol. 24, 6690–6700 (2004).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank G. K. Ojakian for the anti-gp135 antibody, A. Hall, A. Lloyd, R. Y. Tsien, S. Lowe and E. Sahai for constructs, and A. Vaughan for technical assistance with microscopes. We also thank Y. Morishita for discussion on physical forces at cell–cell adhesions. S.D-C. was supported by a FEBS Long Term Fellowship. A.E.P. acknowledges the Interdisciplinary Research Collaboration (IRC) in Nanotechnology (Cambridge, EPSRC UK) and the Dr Mortimer and Theresa Sackler Trust for financial support. This work is supported by MRC funding of the Cell Biology Unit.

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Author notes

  1. Sophie Dupré-Crochet

    Present address: Current address: INSERM UMR 757, Université Paris-sud, Bat 443, 91405 Orsay cedex, France.,

Authors and Affiliations

  1. MRC Laboratory for Molecular Cell Biology and Cell Biology Unit, University College London, Gower Street, WC1E 6BT, London, UK

    Catherine Hogan, Sophie Dupré-Crochet, Mark Norman, Mihoko Kajita, Carola Zimmermann, Franck Pichaud & Yasuyuki Fujita

  2. Department of Cell and Developmental Biology, University College London, Gower Street, WC1E 6BT, London, UK

    Franck Pichaud & Yasuyuki Fujita

  3. London Centre for Nanotechnology and Centre for Nanomedicine, 17–19 Gordon Street, WC1H 0AH, London, UK

    Andrew E. Pelling

  4. Division of Developmental Neurobiology, National Institute for Medical Research, London, NW7 1AA, UK

    Eugenia Piddini, Luis Alberto Baena-López & Jean-Paul Vincent

  5. Imperial College London, The Kennedy Institute of Rheumatology, 1 Aspenlea Road, W6 8LH, London, UK

    Yoshifumi Itoh

  6. Department of Biological Science, Graduate School of Science, Hiroshima University, 739–8526, Higashi-Hiroshima, Japan

    Hiroshi Hosoya

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Contributions

C.H. designed the experiments and generated most of the data; S.D-C. established stable MDCK cell lines and performed statistical analyses (Fig. 1d); M.N. analysed clonal expression of RasV12, RasN17 and RasWT in Drosophila wing imaginal discs (Fig. 1e and Supplementary Information, Fig. S3); M.K. performed western blot analyses (Supplementary Information, Fig. S1b), immunofluorescence studies (Supplementary Information, Fig. S4a) and established stable MDCK cell lines; C.Z. performed western blot and time-lapse analyses (Supplementary Information, Figs S2a, d and S5e), and established stable MDCK cell lines; A.E.P. performed AFM experiments and analyses (Supplementary Information, Fig. S9); E.P., L.A.B-L. and J-P.V. analysed clonal expression of RasV12, RasN17 and RasWT in Drosophila wing imaginal discs (Fig. 1e and Supplementary Information, Fig. S3); Y.I. provided technical expertise on use of collagen; H.H. provided technical expertise on myosin-II; F.P. assisted with Drosophila experiments; Y.F. conceived and designed the study and acted as principal investigator.

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Correspondence to Yasuyuki Fujita.

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The authors declare no competing financial interests.

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Hogan, C., Dupré-Crochet, S., Norman, M. et al. Characterization of the interface between normal and transformed epithelial cells. Nat Cell Biol 11, 460–467 (2009). https://doi.org/10.1038/ncb1853

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