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An siRNA screen identifies the GNAS locus as a driver in 20q amplified breast cancer

Oncogene volume 33, pages 2478–2486 (2014)Cite this article

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Abstract

Poor-prognosis oestrogen receptor-positive breast cancer is characterised by the presence of high-level focal amplifications. We utilised a focused small interfering RNA screen in 14 breast cancer cell lines to define genes that were pathogenic in three genomic regions focally amplified in oestrogen receptor-positive breast cancer, 8p11-12, 11q13 and 20q. Silencing the GNAS locus, that encodes the G protein alpha stimulatory subunit Gαs, specifically reduced the growth of 20q amplified breast cancer cell lines. Examination of a publically available small hairpin RNA data set confirmed GNAS silencing to be selective for 20q amplified cancer cell lines. Cell lines with 20q amplification were found to overexpress specifically the extra long Gαs splice variant (XLαs). Overexpression of XLαs induced cAMP levels to a greater extent than Gαs, suggesting that amplification of the GNAS locus, and overexpression of the XLαs variant in particular, enhanced cAMP signalling. GNAS silencing in amplified cell lines reduced extracellular-signal-regulated kinase 1/2 (ERK1/2) phosphorylation, and conversely overexpression of exogenous XLαs in a non-amplified cell line increased MEK–ERK1/2 phosphorylation, identifying one potential downstream consequence of enhanced cAMP signalling. Our data indicate that amplification of the GNAS locus may contribute to the pathogenesis of breast cancer, and highlight a previously unrecognised role for the GNAS XLαs variant in cancer.

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Acknowledgements

We thank Murat Bastepe for supplying cDNA contructs for Gαs and XLαs. This work was supported by grants from Cancer Research UK and Breakthrough Breast Cancer. Dr Nicholas Turner is a CRUK Clinician Scientist. We acknowledge NHS funding to the NIHR Biomedical Research Centre.

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Authors and Affiliations

  1. The Breakthrough Breast Cancer Research Centre, Institute of Cancer Research, London, UK

    I Garcia-Murillas, R Sharpe, A Pearson, J Campbell, R Natrajan, A Ashworth & N C Turner

  2. Breast Unit, Royal Marsden Hospital, London, UK

    N C Turner

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  1. I Garcia-Murillas
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Correspondence to N C Turner.

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Garcia-Murillas, I., Sharpe, R., Pearson, A. et al. An siRNA screen identifies the GNAS locus as a driver in 20q amplified breast cancer. Oncogene 33, 2478–2486 (2014). https://doi.org/10.1038/onc.2013.202

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