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Transcriptional regulation of the GPX1 gene by TFAP2C and aberrant CpG methylation in human breast cancer

Oncogene volume 32, pages 4043–4051 (2013)Cite this article

Abstract

The complexity of gene regulation has created obstacles to defining mechanisms that establish the patterns of gene expression characteristic of the different clinical phenotypes of breast cancer. TFAP2C is a transcription factor that has a critical role in the regulation of both estrogen receptor-alpha (ERα) and c-ErbB2/HER2 (Her2). Herein, we performed chromatin immunoprecipitation and direct sequencing (ChIP-seq) for TFAP2C in four breast cancer cell lines. Comparing the genomic binding sites for TFAP2C, we identified that glutathione peroxidase (GPX1) is regulated by TFAP2C through an AP-2 regulatory region in the promoter of the GPX1 gene. Knockdown of TFAP2C, but not the related factor TFAP2A, resulted in an abrogation of GPX1 expression. Selenium-dependent GPX activity correlated with endogenous GPX1 expression and overexpression of exogenous GPX1 induced GPX activity and significantly increased resistance to tert-butyl hydroperoxide. Methylation of the CpG island encompassing the AP-2 regulatory region was identified in cell lines where TFAP2C failed to bind the GPX1 promoter and GPX1 expression was unresponsive to TFAP2C. Furthermore, in cell lines where GPX1 promoter methylation was associated with gene silencing, treatment with 5′-aza-2-deoxycytidine (5′-aza-dC) (an inhibitor of DNA methylation) allowed TFAP2C to bind to the GPX1 promoter resulting in the activation of GPX1 RNA and protein expression. Methylation of the GPX1 promoter was identified in ∼20% of primary breast cancers and a highly significant correlation between the TFAP2C and GPX1 expression was confirmed when considering only those tumors with an unmethylated promoter, whereas the related factor, TFAP2A, failed to demonstrate a correlation. The results demonstrate that TFAP2C regulates the expression of GPX1, which influences the redox state and sensitivity to oxidative stress induced by peroxides. Given the established role of GPX1 in breast cancer, the results provide an important mechanism for TFAP2C to further influence oncogenesis and progression of breast carcinoma cells.

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Acknowledgements

We appreciate tumor samples provided by Dr Ryan Askeland and Christine Hochstedler (Department of Pathology, University of Iowa) and statistical support from Dr Junlin Liao (Department of Surgery, University of Iowa). This work was supported in part by the National Institutes of Health grants R01CA109294 (PI: RJ Weigel) and R01CA115438 (PI: FE Domann). Anthony Cyr received salary support from NIH F30 AA019856 and Dr Philip Spanheimer was supported by the NIH grant T32CA148062 (PI: RJ Weigel). The work was also supported by a generous gift from the Kristen Olewine Milke Breast Cancer Research Fund.

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Authors and Affiliations

  1. Department of Surgery, University of Iowa, Iowa City, IA, USA

    M V Kulak, G W Woodfield, M Bogachek, P M Spanheimer & R J Weigel

  2. Department of Radiation Oncology, University of Iowa, Iowa City, IA, USA

    A R Cyr & F E Domann

  3. Department of Biochemistry, University of Iowa, Iowa City, IA, USA

    T Li, D H Price & R J Weigel

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  1. M V Kulak
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Correspondence to R J Weigel.

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Kulak, M., Cyr, A., Woodfield, G. et al. Transcriptional regulation of the GPX1 gene by TFAP2C and aberrant CpG methylation in human breast cancer. Oncogene 32, 4043–4051 (2013). https://doi.org/10.1038/onc.2012.400

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