Article | Published:

A direct role for Met endocytosis in tumorigenesis

Nature Cell Biology volume 13, pages 827837 (2011) | Download Citation

Abstract

Compartmentalization of signals generated by receptor tyrosine kinase (RTK) endocytosis has emerged as a major determinant of various cell functions. Here, using tumour-associated Met-activating mutations, we demonstrate a direct link between endocytosis and tumorigenicity. Met mutants exhibit increased endocytosis/recycling activity and decreased levels of degradation, leading to accumulation on endosomes, activation of the GTPase Rac1, loss of actin stress fibres and increased levels of cell migration. Blocking endocytosis inhibited mutants’ anchorage-independent growth, in vivo tumorigenesis and metastasis while maintaining their activation. One mutant resistant to inhibition by a Met-specific tyrosine kinase inhibitor was sensitive to endocytosis inhibition. Thus, oncogenicity of Met mutants results not only from activation but also from their altered endocytic trafficking, indicating that endosomal signalling may be a crucial mechanism regulating RTK-dependent tumorigenesis.

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Acknowledgements

We thank G. Vande Woude for his gift of the NIH3T3 cell lines expressing murine Met, O. Joffre, V. Carrière, S. Vallath and J. Hulit for their intellectual/technical contributions, A.J. Ridley and S. Heasman for the gift of the GST–PAK-PBD, P. J. Parker and S. Tooze for critically reading the manuscript, J. Camonis, the UK Medical Research Council, and Barts and The London Charitable Foundation.

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Author notes

    • Carine Joffre

    Present address: Institut Curie, INSERM U-830, 26 rue d’Ulm, 75248 Paris Cedex 05, France

Affiliations

  1. Spatial Signalling Team, Centre for Tumour Biology, Barts Cancer Institute, Queen Mary University of London, John Vane Science Centre, Charterhouse Square, London EC1M 6BQ, UK

    • Carine Joffre
    • , Rachel Barrow
    • , Ludovic Ménard
    •  & Stéphanie Kermorgant
  2. Cell Biophysics Laboratory, Cancer Research UK, London Research Institute, 44 Lincoln’s Inn Fields, London WC2A 3PX, UK

    • Véronique Calleja
  3. Centre for Tumour Biology, Barts Cancer Institute, Queen Mary University of London, John Vane Science Centre, Charterhouse Square, London EC1M 6BQ, UK

    • Ian R. Hart

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Contributions

C.J. carried out and analysed most experiments using the NIH3T3 cells both in vitro and in vivo, including characterization of the cell lines, western blots, immunofluorescence analyses, Transwell migration, soft-agar assays, flow cytometry analysis, immunoprecipitation and most separation procedures investigating internalization, degradation and recycling. R.B. did most RNAi knockdowns and transfections followed by the subsequent migration assays, soft-agar assays, western blots and immunofluorescence analysis. R.B. carried out transfection and characterization of hM1268T and hM1268T/N1358H mutants in NIH3T3 cells. L.M. carried out Rac1 activation assays following Grb2 RNAi and PHA treatment, established the detection of active Rac and helped with immunofluorescence analyses and analysis of hM1268T and hM1268T/N1358H cells. V.C. designed and developed the hM1268T and hM1268T/N1358H mutants. S.K. conceived the project, designed experiments, interpreted the data and carried out some of the immunofluorescence and confocal microscopy analyses. I.R.H. advised on the design of the in vivo experiments, trained C.J. and carried out inoculations of tumour cells. S.K. and C.J. wrote the manuscript, with additional input from I.R.H.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Stéphanie Kermorgant.

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https://doi.org/10.1038/ncb2257

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