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Bidirectional cross talk between ERα and EGFR signalling pathways regulates tamoxifen-resistant growth

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Summary

We have previously demonstrated that oestrogen receptor α (ERα) modulates epidermal growth factor receptor (EGFR)/mitogen-activated protein kinase (MAPK) signalling efficiency in a tamoxifen-resistant MCF-7 breast cancer cell line (Tam-R). In the present study we have investigated whether this cross-talk between EGFR/MAPK and ERα signalling pathways is bidirectional by examining the effects of EGFR/MAPK activity on ER functionality in the same cell line. Elevated expression levels of phosphorylated serine 118 (S118) ERα were observed in the Tam-R compared to the parental wild type MCF-7 cell line (WT-MCF-7) under basal growth conditions. Phosphorylation of ERα at S118 was regulated by the EGFR/MAPK pathway in Tam-R cells being increased in response to amphiregulin (AR) and inhibited by the selective EGFR tyrosine kinase inhibitor, gefitinib and the MEK1/2 inhibitor, PD184352. Recruitment of the co-activators p68 RNA helicase and SRC1 to ERα, oestrogen response element (ERE) activity and Tam-R cell growth were similarly EGFR/MAPK-regulated. Chromatin immunoprecipitation (ChIP) studies revealed that in Tam-R cells the ERα assembled on the AR gene promoter and this was associated with elevated basal expression of AR mRNA. Furthermore, AR mRNA expression was under the regulation of the EGFR/MAPK and ERα signalling pathways. Neutralising antibodies to AR inhibited EGFR/ERK1/2 activity, reduced S118 ERα phosphorylation and reduced AR mRNA expression in TAM-R cells. These findings suggest that ERα function in Tam-R cells is maintained as a consequence of EGFR/MAPK-mediated phosphorylation at serine residue 118 resulting in the generation of a self-propogating autocrine growth-regulatory loop through the ERα-mediated production of AR.

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Acknowledgements

The authors would like to thank Drs Kari Rossow and Ralf Janknecht of the Department of Biochemistry and Molecular Biology, Mayo Clinic, USA for the kind gift of the p68 RNA Helicase antibody and Dr. Judith Sebolt-Leopold for kindly providing the MEK1/2 inhibitor PD184352. We would also like to thank Carol Dutkowski and Pauline Finlay for expert technical assistance and Lynne Farrow for performing the statistical analyses. This research was generously supported by the Tenovus organisation.

‘Faslodex’ and ‘IRESSA’ are trademarks of the AstraZeneca group of companies

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Authors and Affiliations

  1. Tenovus Centre for Cancer Research, Welsh School of Pharmacy, Cardiff University, Cardiff, UK

    D.J. Britton, J.M. Knowlden, D. Barrow, M. Giles, R.A. McClelland, J.M.W. Gee & R.I. Nicholson

  2. Tenovus Centre for Cancer Research, Welsh School of Pharmacy, Cardiff University, Redwood Building, King Edward VII Avenue, CF10 3XF, Cardiff, UK

    I.R. Hutcheson

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  1. D.J. Britton
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Britton, D., Hutcheson, I., Knowlden, J. et al. Bidirectional cross talk between ERα and EGFR signalling pathways regulates tamoxifen-resistant growth. Breast Cancer Res Treat 96, 131–146 (2006). https://doi.org/10.1007/s10549-005-9070-2

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