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Hsp-27 induction requires POU4F2/Brn-3b TF in doxorubicin-treated breast cancer cells, whereas phosphorylation alters its cellular localisation following drug treatment

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Cell Stress and Chaperones Aims and scope

Abstract

POU4F2/Brn-3b transcription factor (referred to as Brn-3b) is elevated in >60% of breast cancers and profoundly alters growth and behaviour of cancer cells by regulating distinct subsets of target genes. Previous studies showed that Brn-3b was required to maximally transactivate small heat shock protein, HSPB1/Hsp-27 (referred to as Hsp-27), and consequently, Brn-3b expression correlated well with Hsp27 levels in human breast biopsies. In these studies, we showed that Brn-3b is increased in MCF7 breast cancer cells that survive following treatment with chemotherapeutic drug doxorubicin (Dox) with concomitant increases in Hsp-27 expression. Targeting of Brn-3b using short interfering RNA reduced Hsp-27 in Dox-treated cells, suggesting that Brn-3b regulates Hsp-27 expression under these conditions. Wound healing assays showed increased Brn-3b in Dox-treated migratory cells that also express Hsp-27. Interestingly, Hsp-27 phosphorylation and cellular localisation are also significantly altered at different times following Dox treatment. Thus, phospho-Hsp-27 (p-Hsp27) protein displayed widespread distribution after 24 hrs of Dox treatment but was restricted to the nucleus after 5 days. However, in drug-resistant cells (grown in Dox for > 1 month), p-Hsp-27 was excluded from nuclei and most of the cytoplasm and appeared to be associated with the cell membrane. Studies to determine how this protein promotes survival and migration in breast cancer cells showed that the protective effects were conferred by unphosphorylated Hsp-27 protein. Thus, complex and dynamic mechanisms underlie effects of Hsp-27 protein in breast cancer cells following treatment with chemotherapeutic drugs such as Dox, and this may contribute to invasiveness and drug resistance following chemotherapy.

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Abbreviations

Brn-3b:

POU transcription factorBrn-3b/POU4F2

HSP:

Small heat-shock protein

AV:

Adenovirus

WT:

Wild type

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

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Acknowledgements

This work was supported by British Heart Foundation (BHF), Breast Cancer Campaign (BCC) UK and Association for International Cancer Research (AICR), UK. We also thank Dr. J.L. Martin, Department of Physiology, Cardiovascular Institute, Loyola University, Chicago, IL, USA, for sharing Hsp-27 wild-type and mutant adenoviral constructs and Dr. D. Trott and Prof M. Rose, Imperial College London, for providing viral stocks, which were invaluable for these experiments.

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  1. Medical Molecular Biology Unit, University College London, 30 Guilford Street, London, WC1N 1EH, UK

    Rieko Fujita, Samir Ounzain, Alice Chun Yin Wang & Vishwanie Shanie Budhram-Mahadeo

  2. Cardiovascular Division, Kings College London, Department of Cardiology, The Rayne Institute, St Thomas’s Hospital, Lambeth Palace Road, SE1 7EH, London, UK

    Richard John Heads

  3. Cardiology Department, The Rayne Institute, St Thomas’s Hospital, Lambeth Palace Road, London, SE1 7EH, UK

    Richard John Heads

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  1. Rieko Fujita
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Correspondence to Vishwanie Shanie Budhram-Mahadeo.

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Fujita, R., Ounzain, S., Wang, A.C.Y. et al. Hsp-27 induction requires POU4F2/Brn-3b TF in doxorubicin-treated breast cancer cells, whereas phosphorylation alters its cellular localisation following drug treatment. Cell Stress and Chaperones 16, 427–439 (2011). https://doi.org/10.1007/s12192-011-0256-8

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