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The Forkhead Box M1 protein regulates BRIP1 expression and DNA damage repair in epirubicin treatment

Oncogene volume 32, pages 4634–4645 (2013)Cite this article

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Abstract

FOXM1 is implicated in genotoxic drug resistance but its role and mechanism of action remain unclear. Here, we establish that γH2AX foci, indicative of DNA double-strand breaks (DSBs), accumulate in a time-dependent manner in the drug-sensitive MCF-7 cells but not in the resistant counterparts in response to epirubicin. We find that FOXM1 expression is associated with epirubicin sensitivity and DSB repair. Ectopic expression of FOXM1 can increase cell viability and abrogate DSBs sustained by MCF-7 cells following epirubicin, owing to an enhancement in repair efficiency. Conversely, alkaline comet and γH2AX foci formation assays show that Foxm1-null cells are hypersensitive to DNA damage, epirubicin and γ-irradiation. Furthermore, we find that FOXM1 is required for DNA repair by homologous recombination (HR) but not non-homologous end joining (NHEJ), using HeLa cell lines harbouring an integrated direct repeat green fluorescent protein reporter for DSB repair. We also identify BRIP1 as a direct transcription target of FOXM1 by promoter analysis and chromatin-immunoprecipitation assay. In agreement, depletion of FOXM1 expression by small interfering RNA downregulates BRIP1 expression at the protein and mRNA levels in MCF-7 and the epirubicin-resistant MCF-7 EpiR cells. Remarkably, the requirement for FOXM1 for DSB repair can be circumvented by reintroduction of BRIP1, suggesting that BRIP1 is an important target of FOXM1 in DSB repair. Indeed, like FOXM1, BRIP1 is needed for HR. These data suggest that FOXM1 regulates BRIP1 expression to modulate epirubicin-induced DNA damage repair and drug resistance.

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Acknowledgements

EW-FL, ARG and RCC were supported by grants from Cancer Research UK, EW-FL and C-YK by grants from Breast Cancer Campaign and Imperial College NHS Trust, LJM grants from Fundação para a Ciência e a Tecnologia, and PK and MK from the Royal Thai Government Scholarships. We also thank Dr Keng Heng for help with the microscopy and flow cytometry work.

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Authors and Affiliations

  1. Department of Surgery and Cancer, Imperial College London, Hammersmith Hospital Campus, London, UK

    L J Monteiro, P Khongkow, M Kongsema, C-Y Koo, A R Gomes, P H Pinto, V Varghese, L M Kenny, R Charles Coombes & E W-F Lam

  2. School of Cancer Sciences, College of Medical and Dental Sciences, University of Birmingham, Edgbaston, Birmingham, UK

    J R Morris

  3. Department of Applied Biology and Chemical Technology, The Hong Kong Polytechnic University, Hung Hom, Kowloon, Hong Kong

    C Man

  4. Department of Medical and Molecular Genetics, King’s College London, Guy’s Hospital, London, UK

    D Weekes

  5. Unidad de Investigación, Hospital Universitario de Canarias, Instituto de Tecnologías Biomédicas, Ofra s/n, La Laguna, Tenerife, Spain

    R Freire

  6. Division of Cell Biology, The Netherlands Cancer Institute, Amsterdam, CX, The Netherlands

    R H Medema

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Monteiro, L., Khongkow, P., Kongsema, M. et al. The Forkhead Box M1 protein regulates BRIP1 expression and DNA damage repair in epirubicin treatment. Oncogene 32, 4634–4645 (2013). https://doi.org/10.1038/onc.2012.491

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