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Functional identification of genes causing estrogen independence of human breast cancer cells

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Abstract

Endocrine treatment of breast cancer is widely applied and effective. However, in advanced disease cases, the tumors will eventually progress into an estrogen-independent and therapy-resistant phenotype. To elucidate the molecular mechanisms underlying this endocrine therapy failure, we applied retroviral insertion mutagenesis to identify the main genes conferring estrogen independence to human breast cancer cells. Estrogen-dependent ZR-75-1 cells were infected with replication-defective retroviruses followed by selection with the anti-estrogen 4-hydroxy-tamoxifen. In the resulting panel of 79 tamoxifen-resistant cell lines, the viral integrations were mapped within the human genome. Genes located in the immediate proximity of the retroviral integration sites were characterized for altered expression and their capacity to confer anti-estrogen resistance when transfected into breast cancer cells. Out of 15 candidate BCAR (breast cancer anti-estrogen resistance) genes, seven (AKT1, AKT2, BCAR1, BCAR3, EGFR, GRB7, and TRERF1/BCAR2) were shown to directly underlie estrogen independence. Our results show that insertion mutagenesis is a powerful tool to identify BCAR loci, which may provide insights into the molecular and cellular mechanisms of breast tumor progression and therapy resistance thereby offering novel targets for the development of tailor-made therapeutical and prevention strategies.

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Acknowledgements

We acknowledge Anton Berns (Amsterdam) for the splinkerette protocol, and Fred Sweep and Nicolai Grebenchtchikov (Nijmegen) for their support in generating antibodies. Excellent support of various current and former members of our research laboratory is gratefully acknowledged. For stimulating discussions and support, we are indebted to Ad Brinkman, Danielle Meijer, John Foekens, Maxime Look, Leendert Looijenga, Els Berns, John Martens, Maurice Jansen and Riccardo Fodde. Funding: This study was supported by grants of the Dutch Cancer Society (DDHK96-1245 & 99-1883), the Susan G. Komen Breast Cancer Foundation (BCTR0100675), the Association for International Cancer Research (04–148) and the Josephine Nefkens Stichting.

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Authors and Affiliations

  1. Department of Pathology, Josephine Nefkens Institute, Be 432, Erasmus MC-University Medical Center Rotterdam, P.O. Box 2040, Rotterdam, CA, 3000, The Netherlands

    Ton van Agthoven, Jos Veldscholte, Marcel Smid, Thecla L. A. van Agthoven, Lilian Vreede, Marieke Broertjes, Ingrid de Vries, Danielle de Jong, Roya Sarwari & Lambert C. J. Dorssers

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  1. Ton van Agthoven
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  2. Jos Veldscholte
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  10. Lambert C. J. Dorssers
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Correspondence to Lambert C. J. Dorssers.

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Ton van Agthoven and Jos Veldscholte contributed equally to this research.

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van Agthoven, T., Veldscholte, J., Smid, M. et al. Functional identification of genes causing estrogen independence of human breast cancer cells. Breast Cancer Res Treat 114, 23–30 (2009). https://doi.org/10.1007/s10549-008-9969-5

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