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Localized and reversible TGFβ signalling switches breast cancer cells from cohesive to single cell motility

Nature Cell Biology volume 11, pages 1287–1296 (2009)Cite this article

Abstract

Here we use intravital imaging to demonstrate a reversible transition to a motile state as breast cancer cells spread. Imaging primary tumours revealed heterogeneity in cell morphology and motility. Two distinct modes of motility were observed: collective and single-celled. By monitoring the localization of Smad2 and the activity of a TGFβ-dependent reporter gene during breast cancer cell dissemination, we demonstrate that TGFβ signalling is transiently and locally activated in motile single cells. TGFβ1 switches cells from cohesive to single cell motility through a transcriptional program involving Smad4, EGFR, Nedd9, M-RIP, FARP and RhoC. Blockade of TGFβ signalling prevented cells moving singly in vivo but did not inhibit cells moving collectively. Cells restricted to collective invasion were capable of lymphatic invasion but not blood-borne metastasis. Constitutive TGFβ signalling promoted single cell motility and intravasation but reduced subsequent growth in the lungs. Thus, transient TGFβ signalling is essential for blood-borne metastasis.

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Figure 1: Transient acquisition of motile behaviour by breast cancer cells.
Figure 2: TGFβ signalling in MTLn3E cells.
Figure 3: Nuclear accumulation of Smad2 in singly-moving cells.
Figure 4: Activation of Smad-dependent transcription in singly moving cells.
Figure 5: TGFβ requires Smad4 to switch cells to single cell motility.
Figure 6: Roles of multiple TGFβ target genes in the switch to single cell motility.
Figure 7: TGFβ signalling is required for single cell motility in vivo.
Figure 8: TGFβ signalling is required for haematogenous but not lymphatic metastasis.

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Acknowledgements

We thank lab colleagues for advice and comments on this project. We thank members of the LRI FACS laboratory, Clare Watkins and members of the LRI Biological Resources Unit, the Paterson Institute for Cancer Research microarray facility, the LRI Bioinformatics team, Mike Howell of the LRI Developmental Signalling laboratory and Debbie Aubyn of the LRI microscopy laboratory for technical assistance. This work was funded by the Breast Cancer Campaign project grant 12May05 and CRUK.

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Authors and Affiliations

  1. Tumour Cell Biology Laboratory, CR-UK London Research Institute, 44 Lincoln's Inn Fields, London, WC2A 3PX, UK

    Silvia Giampieri, Cerys Manning, Steven Hooper & Erik Sahai

  2. Centre for Tumour Biology, Institute of Cancer, Charterhouse Square, London, EC1M 6BQ, UK

    Louise Jones

  3. Developmental Signalling Laboratory, CR-UK London Research Institute, 44 Lincoln's Inn Fields, London, WC2A 3PX, UK

    Caroline S. Hill

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  1. Silvia Giampieri
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Contributions

S.G. generated most of the data in Figs 2, 3, 4, 5 and 7 with some assistance from E.S. S.G. and E.S. made equal contributions to Fig. 8 with assistance from C.S.M.; E.S. generated most of the data in Figs 1 and 6 with assistance from S.G. and S.H.; L.J.J. provided clinical material used in Supplementary Information figures; S.G. generated data shown in Supplementary Information Figs 3, 4, 5, 6, 7, 8; S.H. and C.S.M. assisted with Supplementary Information Figs 7 and 8, respectively; E.S. generated data in Supplementary Information Figs 1, 2, 6 and 9; C.S.H. provided reagents, expertise and intellectual input. S.G. and E.S. designed the experiments.

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Correspondence to Erik Sahai.

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The authors declare no competing financial interests.

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Giampieri, S., Manning, C., Hooper, S. et al. Localized and reversible TGFβ signalling switches breast cancer cells from cohesive to single cell motility. Nat Cell Biol 11, 1287–1296 (2009). https://doi.org/10.1038/ncb1973

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