Abstract
Genetic changes in HER2, PTEN, PIK3CA and AKT1 are all common in breast cancer and lead to the elevated phosphorylation of downstream targets of the PI3K/AKT signalling pathway. Changes in HER2, PTEN, PIK3CA and AKT have all been reported to lead to both enhanced proliferation and failures in hollow lumen formation in three dimensional epithelial culture models, but it is not clear whether these failures in lumen formation are caused by any failure in the spatial coordination of lumen formation (hollowing) or purely a failure in the apoptosis and clearance of luminal cells (cavitation). Here, we use normal murine mammary gland (NMuMG) epithelial cells, which form a hollow lumen without significant apoptosis, to compare the transformation by these four genetic changes. We find that either mutant PIK3CA expression or PTEN loss, but not mutant AKT1 E17K, cause disrupted epithelial architecture, whereas HER2 overexpression drives strong proliferation without affecting lumen formation in these cells. We also show that PTEN requires both lipid and protein phosphatase activity, its extreme C-terminal PDZ binding sequence and probably Myosin 5A to control lumen formation through a mechanism that does not correlate with its ability to control AKT, but which is selectively lost through mutation in some tumours. These findings correlate AKT-independent signalling activated by mutant PIK3CA or PTEN loss, but not strongly by HER2, with disrupted epithelial architecture and tumour formation.
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Acknowledgements
We thank Jon Backer (Albert Einstein College of Medicine) and Dario Alessi, David Meek, Francis Fuller-Pace, Virginia Appleyard, Alastair Thompson, Emily Davis, John Rouse, Inke Näthke and Paul Crocker (all University of Dundee) for reagents and Peter Downes (University of Dundee) for helpful discussions. We thank Sam Swift, Paul Appleton, Martin Kierans and John James (University of Dundee, CHIPS microscopy facility) for assistance with light and electron microscopy. FB and JCL have been funded by a project grant from the Association for International Cancer Research and NRW is a Wellcome Trust Prize Student. Work in the Inositol Lipid Signalling laboratory is funded by the Medical Research Council, the Association for International Cancer Research and the pharmaceutical companies of the DSTT consortium (Astra Zeneca, Boehringer Ingelheim, GlaxoSmithKline, Merck Serono and Pfizer). We wish to dedicate this work to the memory of Joseph Lim, a valued colleague and friend.
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Research in the Inositol Lipid Signalling laboratory has been partly funded by the pharmaceutical companies of the DSTT consortium (Astra Zeneca, Boehringer Ingelheim, GlaxoSmithKline, Merck Serono and Pfizer).
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Berglund, F., Weerasinghe, N., Davidson, L. et al. Disruption of epithelial architecture caused by loss of PTEN or by oncogenic mutant p110α/PIK3CA but not by HER2 or mutant AKT1. Oncogene 32, 4417–4426 (2013). https://doi.org/10.1038/onc.2012.459
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DOI: https://doi.org/10.1038/onc.2012.459
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