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Competition amongst Eph receptors regulates contact inhibition of locomotion and invasiveness in prostate cancer cells

Nature Cell Biology volume 12, pages 1194–1204 (2010)Cite this article

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Abstract

Metastatic cancer cells typically fail to halt migration on contact with non-cancer cells. This invasiveness is in contrast to normal mesenchymal cells that retract on contact with another cell. Why cancer cells are defective in contact inhibition of locomotion is not understood. Here, we analyse the dynamics of prostate cancer cell lines co-cultured with fibroblasts, and demonstrate that a combinatorial code of Eph receptor activation dictates whether cell migration will be contact inhibited. The unimpeded migration of metastatic PC-3 cells towards fibroblasts is dependent on activation of EphB3 and EphB4 by ephrin-B2, which we show activates Cdc42 and cell migration. Knockdown of EphB3 and EphB4 restores contact inhibition of locomotion to PC-3 cells. Conversely, homotypic collisions between two cancer cells results in contact inhibition of locomotion, mediated by EphA–Rho–Rho kinase (ROCK) signalling. Thus, the migration of cancer cells can switch from restrained to invasive, depending on the Eph-receptor profile of the cancer cell and the reciprocal ephrin ligands expressed by neighbouring cells.

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Figure 1: Failure of CIL by PC-3 cells on contact with fibroblasts.
Figure 2: Ephrin-A ligands are sufficient to induce CIL between PC-3 cells.
Figure 3: EphA2 and EphA4 are required for CIL between PC-3 cells.
Figure 4: Ephrin-B2 stimulates PC-3 cell migration.
Figure 5: Ephrin-B2 induces filopodia by activating Cdc42 in PC-3 cells.
Figure 6: EphB3 and EphB4 knockdown restores CIL of PC-3 cells on contact with fibroblasts.
Figure 7: Immunohistochemical staining of EphB4 and ephrin-B2 in prostate cancer.
Figure 8: Model of CIL regulation in PC-3 cells.

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  • 19 November 2010

    In the version of this article initially published online, Fig. 4c was incorrectly labelled on the y axis. This error has been corrected in both the HTML and PDF versions of the article.

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Acknowledgements

We thank H. Ross for technical assistance and I. Hers for advice on immunoprecipitations, M. Brown for BMECs, A. Ziemiecki and A. -C. Andres for anti-EphB4, D. Wilkinson for anti-EphA4 and E. Pasquale for anti-EphB3. We thank P. Martin, N. Perkins and C. Paraskeva for critically reading the manuscript. We are grateful to J. Dovovan and NCRI/MRC ProMPT collaborative for facilitating clinical aspects of the work. This study has been approved by the Southmead Research Ethics Committee and was funded by a Wellcome Trust PhD studentship to J.B., an MRC studentship to S.K. and a Cancer Research UK project grant to C.D.N.

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Authors and Affiliations

  1. School of Biochemistry, University of Bristol, Bristol, BS8 1TD, UK

    Jonathan W. Astin, Jennifer Batson, Shereen Kadir, Jessica Charlet & Catherine D. Nobes

  2. School of Physiology and Pharmacology, University of Bristol, Bristol, BS8 1TD, UK

    Catherine D. Nobes

  3. Bristol Urological Institute, Southmead Hospital, Bristol, BS10 5NB, UK

    Raj A. Persad & David Gillatt

  4. Department of Cellular Pathology, Southmead Hospital, Bristol, BS10 5NB, UK

    Jon D. Oxley

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Contributions

J.W.A., S.K. and C.D.N. designed experiments. J.W.A. and J.C. performed the RT–PCR, J.B. performed the Cdc42-knockdown experiments, C.D.N. performed microinjection experiments and immunohistochemistry, and J.W.A. carried out all other experiments. D.G. and R.P. provided prostate tissue and J.O., J.W.A. and C.D.N. prepared and analysed prostate immunohistochemistry. J.W.A. and C.D.N. wrote the manuscript.

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Correspondence to Catherine D. Nobes.

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Astin, J., Batson, J., Kadir, S. et al. Competition amongst Eph receptors regulates contact inhibition of locomotion and invasiveness in prostate cancer cells. Nat Cell Biol 12, 1194–1204 (2010). https://doi.org/10.1038/ncb2122

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