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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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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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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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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DOI: https://doi.org/10.1038/ncb1973
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