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Cellular and Molecular Biology

Targeting peroxiredoxin 1 impairs growth of breast cancer cells and potently sensitises these cells to prooxidant agents

British Journal of Cancervolume 119pages873884 (2018) | Download Citation



Our previous work has shown peroxiredoxin-1 (PRDX1), one of major antioxidant enzymes, to be a biomarker in human breast cancer. Hereby, we further investigate the role of PRDX1, compared to its close homolog PRDX2, in mammary malignant cells.


CRISPR/Cas9- or RNAi-based methods were used for genetic targeting PRDX1/2. Cell growth was assessed by crystal violet, EdU incorporation or colony formation assays. In vivo growth was assessed by a xenotransplantation model. Adenanthin was used to inhibit the thioredoxin-dependent antioxidant defense system. The prooxidant agents used were hydrogen peroxide, glucose oxidase and sodium L-ascorbate. A PY1 probe or HyPer-3 biosensor were used to detect hydrogen peroxide content in samples.


PRDX1 downregulation significantly impaired the growth rate of MCF-7 and ZR-75-1 breast cancer cells. Likewise, xenotransplanted PRDX1-deficient MCF-7 cells presented a retarded tumour growth. Furthermore, genetic targeting of PRDX1 or adenanthin, but not PRDX2, potently sensitised all six cancer cell lines studied, but not the non-cancerous cells, to glucose oxidase and ascorbate.


Our study pinpoints the dominant role for PRDX1 in management of exogeneous oxidative stress by breast cancer cells and substantiates further exploration of PRDX1 as a target in this disease, especially when combined with prooxidant agents.

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Note: This work is published under the standard license to publish agreement. After 12 months the work will become freely available and the license terms will switch to a Creative Commons Attribution 4.0 International (CC BY 4.0).


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The authors wish to thank Prof. Rafal Ploski and Dr. Malgorzata Rydzanicz (both of Medical University of Warsaw) for their invaluable contribution to RNASeq studies. We also thank Dr. Beata Pyrzynska (Medical University of Warsaw) for her help in obtaining HEK-293FT cells. We would like to thank Dr. Rut Klinger (University College Dublin) for her help in generating the HyPer3-encoding lentiviral vector. We want to thank Mr. Tomasz Jarzynka (International Institute of Molecular and Cell Biology in Warsaw) for help with script writing for ImageJ automated analysis. We also want to thank Prof. Pawel Wlodarski and Dr. Ilona Kalaszczynska (both of Medical University of Warsaw) for providing an access to Nikon microscope.

Authors contributions

M.B., A.O.Z. and A.M. cultured the cells; A.O.Z. and M.B. carried out the cell growth/survival assays and western blotting experiments; M.B. performed flow cytometry-based study; A.G.-J. performed the experiments with PY1 probe; A.D. and A.G.-J. carried out digital analysis of the colony formation images; A.O.Z., M.B. and A.D. carried out selection of sgRNA clones for the in vitro study; A.M. generated the MCF-7 cells for the in vivo study; R.Z., A.M., M.F., A.O.Z., M.B., A.G.-J. and M.S. carried out the lentiviral transduction procedures; A.M. and A.Z. carried out the in vivo study; M.F. generated the MCF-7sgRNA derivatives for the in vivo study and oversaw the in vivo study; P.S. carried out the in silico analysis of ADNT:PRDX interactions; M.B. and A.O.Z. carried out life imaging studies; P.G. analysed the clinical data; L.T. carried out the RNAseq study; P.C.O'.L. generated the sgRNA sequences and consulted the CRISPR/Cas9-mediated generation of MCF-7 cell line derivatives; M.K. and M.B. carried out the Cs-137-irradiation study; M.B. carried out the statistical analysis; M.B. and R.Z. designed the experiments and drafted the manuscript; J.G. provided critical insights into the study design, provided research materials for oxidative stress assessment, and contributed to supervision of the study; and R.Z. conceived the study, supervised the project, and coordinated all experiments in this work. All authors critically revised the initial draft of the manuscript and subsequent revisions. All authors approved the manuscript in its current form.

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Author notes

  1. These authors contributed equally: Malgorzata Bajor and Agata O. Zych


  1. Department of Immunology, Center of Biostructure Research, Medical University of Warsaw, Warsaw, Poland

    • Malgorzata Bajor
    • , Agata O. Zych
    • , Agnieszka Graczyk-Jarzynka
    • , Angelika Muchowicz
    • , Malgorzata Firczuk
    • , Antoni Domagala
    • , Marta Siernicka
    • , Agnieszka Zagozdzon
    • , Jakub Golab
    •  & Radoslaw Zagozdzon
  2. Department of Clinical Immunology, Transplantation Institute, Medical University of Warsaw, Warsaw, Poland

    • Malgorzata Bajor
    • , Monika Kniotek
    •  & Radoslaw Zagozdzon
  3. Postgraduate School of Molecular Medicine, Medical University of Warsaw, Warsaw, Poland

    • Agata O. Zych
    •  & Marta Siernicka
  4. Department of Medical Genetics, Medical University of Warsaw, Warsaw, Poland

    • Lech Trzeciak
  5. Center for Advanced Materials and Technologies, Warsaw University of Technology, Warsaw, Poland

    • Lech Trzeciak
  6. Laboratory of Human Cancer Genetics, Centre of New Technologies, University of Warsaw, Warsaw, Poland

    • Pawel Gaj
  7. Department of Bioinformatics, Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland

    • Pawel Siedlecki
    •  & Radoslaw Zagozdzon
  8. Department of Systems Biology, Institute of Experimental Plant Biology and Biotechnology, University of Warsaw, Warsaw, Poland

    • Pawel Siedlecki
  9. Helen Diller Family Comprehensive Cancer Center, University of California San Francisco, San Francisco, CA, USA

    • Patrick C. O’Leary
  10. Centre for Preclinical Research and Technology, Medical University of Warsaw, Warsaw, Poland

    • Jakub Golab


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Ethics approval and consent to participate

All animal experiments were performed in accordance with the guidelines approved by the Ethics Committee of the Medical University of Warsaw (approval No. 40/2015).


The work was supported by National Science Centre, Poland (grant No. 2014/13/B/NZ5/01354; RZ), European Commission Horizon 2020 Programme (692180-STREAM-H2020-TWINN-2015; J.G.), European Commission 7th Framework Programme (FP7-REGPOT-2012-CT2012-316254-BASTION; J.G.), and Medical University of Warsaw (1M19/PM14/14; M.B.).

Competing interests

The authors declare no competing interests.


This work is published under the standard license to publish agreement. After 12 months the work will become freely available and the license terms will switch to a Creative Commons Attribution-NonCommercial-Share Alike 4.0 Unported License).

Corresponding author

Correspondence to Radoslaw Zagozdzon.

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