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Decreased levels of hypoxic cells in gefitinib treated ER+ HER-2 overexpressing MCF-7 breast cancer tumors are associated with hyperactivation of the mTOR pathway: therapeutic implications for combination therapy with rapamycin

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Abstract

Developing novel synergistic and more effective combination treatments is necessary for better management of breast cancer in the clinic. It is established that HER-2 overexpressing breast cancers are sensitive to the HER-1 (epidermal growth factor receptor (EGFR)) inhibitor gefitinib, but that this targeted agent produces only moderate therapeutic effects in vivo. Here, we use a model of ER+ HER-2 overexpressing MCF-7 breast cancer (MCF-7HER-2) to identify, as broadly as possible, the in vivo microenvironmental and molecular therapeutic responses to gefitinib to predict a therapeutically viable target for gefitinib-based combination treatment. Our data show a link between in vivo reductions in tumor hypoxia (3-fold decrease, P = 0.002) and elevated activity of the mTOR pathway (3.8-fold increase in phospho-p70-S6K protein, P = 0.006) in gefitinib treated MCF-7HER-2 tumors. Despite decreased levels of phosphorylated EGFR, HER-2 and Erk1/2 (P = 0.081, 0.005 and 0.034, respectively) the expression of phospho-AKT was not reduced in MCF-7HER-2 tumors after gefitinib treatment. Levels of ERα receptor were, however, 1.8-fold higher in gefitinib treated compared to control tumors (P = 0.008). Based on these results we predict that gefitinib activity against ER+ HER-2 overexpressing EGFR co-expressing breast cancers should be enhanced if used with agents that target the mTOR pathway. In vitro studies using MCF-7HER-2 and BT474 breast cancer cells exposed to gefitinib and rapamycin in combination show that this combination produced significantly greater growth inhibitory effects than either of the drugs alone. Chou and Talalay analysis of the data suggested that combination of gefitinib and rapamycin was synergistic (CI < 1) at a number of selected drug ratios and over a broad range of effective doses.

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Notes

  1. Personal communication: Dr. Karen Gelmon; Medical Oncology, British Columbia Cancer Agency, Vancouver, British Columbia, Canada.

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Acknowledgements

The authors thank Drs. Sandra Dunn, Sylvia Ng, Dawn Waterhouse, Ghania Chikh and members of the Advanced Therapeutics Laboratory for helpful comments. We also thank Dr. Gerry Krystal and Dr. Sandra Dunn for sharing their antibody stock. We greatly appreciate the help of Tom Oliver with western blotting, Hong Yan with cytotoxicity assays and Dana Masin, Rebecca Ng, and Maryam Osooly with animal work.

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Authors and Affiliations

  1. Department of Advanced Therapeutics, British Columbia Cancer Agency, 675 West 10th Ave., Vancouver, BC, Canada, V5Z 1L3

    Wieslawa H. Dragowska, Maïté Verreault, Donald T. T. Yapp, Corinna Warburton, Lincoln Edwards, Euan C. Ramsay & Marcel B. Bally

  2. Department of Medical Biophysics, British Columbia Cancer Research Centre, Vancouver, BC, Canada

    Lynsey A. Huxham & Andrew I. Minchinton

  3. Faculty of Pharmaceutical Sciences ,The Division of Pharmaceutics and Biopharmaceutics, University of British Columbia, Vancouver, BC, Canada

    Donald T. T. Yapp

  4. Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada

    Andrew I. Minchinton & Marcel B. Bally

  5. Faculty of Pharmaceutical Sciences, University of British Columbia, Vancouver, BC, Canada

    Marcel B. Bally

  6. Medical Oncology, British Columbia Cancer Agency, Vancouver, BC, Canada

    Karen Gelmon

Authors
  1. Wieslawa H. Dragowska
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  2. Maïté Verreault
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  7. Lynsey A. Huxham
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  8. Andrew I. Minchinton
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  9. Karen Gelmon
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  10. Marcel B. Bally
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Correspondence to Wieslawa H. Dragowska.

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This work was supported by the Canadian Breast Cancer Research Alliance in association with the National Cancer Institute of Canada and the Canadian Institutes of Health Research.

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Dragowska, W.H., Verreault, M., Yapp, D.T.T. et al. Decreased levels of hypoxic cells in gefitinib treated ER+ HER-2 overexpressing MCF-7 breast cancer tumors are associated with hyperactivation of the mTOR pathway: therapeutic implications for combination therapy with rapamycin. Breast Cancer Res Treat 106, 319–331 (2007). https://doi.org/10.1007/s10549-007-9502-2

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