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Roles of Atox1 and p53 in the trafficking of copper-64 to tumor cell nuclei: implications for cancer therapy

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Abstract

Owing to its cytotoxicity, free copper is chelated by protein side chains and does not exist in vivo. Several chaperones transport copper to various cell compartments, but none have been identified that traffic copper to the nucleus. Copper-64 decays by β + and β − emission, allowing positron emission tomography and targeted radionuclide therapy for cancer. Because the delivery of 64Cu to the cell nucleus may enhance the therapeutic effect of copper radiopharmaceuticals, elucidation of the pathway(s) involved in transporting copper to the tumor cell nucleus is important for optimizing treatment. We identified Atox1 as one of the proteins that binds copper in the nucleus. Mouse embryonic fibroblast cells, positive and negative for Atox1, were used to determine the role of Atox1 in 64Cu transport to the nucleus. Mouse embryonic fibroblast Atox1+/+ cells accumulated more 64Cu in the nucleus than did Atox1−/− cells. HCT 116 colorectal cancer cells expressing p53 (+/+) and not expressing p53 (−/−) were used to evaluate the role of this tumor suppressor protein in 64Cu transport. In cells treated with cisplatin, the uptake of 64Cu in the nucleus of HCT 116 p53+/+ cells was greater than that in HCT 116 p53−/− cells. Atox1 expression increased in HCT 116 p53+/+ and p53−/− cells treated with cisplatin; however, Atox1 localized to the nuclei of p53+/+ cells more than in the p53−/− cells. The data presented here indicate that Atox1 is involved in copper transport to the nucleus, and cisplatin affects nuclear transport of 64Cu in HCT 116 cells by upregulating the expression and the nuclear localization of Atox1.

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Abbreviations

BSA:

Bovine serum albumin

Ctr1:

Copper transporter 1

HPLC:

High-performance liquid chromatography

mAb:

Monoclonal antibody

MEF:

Mouse embryonic fibroblast

PBS:

Phosphate-buffered saline

PET:

Positron emission tomography

siRNA:

Small interfering RNA

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Acknowledgments

This research was supported by National Institutes of Health/National Cancer Institute grant 5R01CA064475 (C.J.A.) and Department of Energy grant DE-FG02-08ER64671 (Integrated Research Training Program of Excellence in Radiochemistry awarded to Suzanne Lapi supporting Y.G.). The mass spectrometry portion of the project was supported by grants from the National Center for Research Resources (5P41RR000954-35) and the National Institute of General Medical Sciences (8 P41 GM103422-35) from the National Institutes of Health.

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Author notes

  1. Albert J. Chang

    Present address: Department of Radiation Oncology, University of California, San Francisco, 1600 Divisadero Street, Suite H1031, Box 1708, San Francisco, CA, 94115, USA

Authors and Affiliations

  1. Department of Radiology, University of Pittsburgh, 100 Technology Drive, Suite 452, Pittsburgh, PA, 15219, USA

    Wissam Beaino & Carolyn J. Anderson

  2. Department of Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, PA, USA

    Carolyn J. Anderson

  3. Department of Bioengineering, University of Pittsburgh, Pittsburgh, PA, USA

    Carolyn J. Anderson

  4. Mallinckrodt Institute of Radiology, Washington University School of Medicine, St. Louis, MO, USA

    Yunjun Guo

  5. Department of Radiation Oncology, Washington University School of Medicine, St. Louis, MO, USA

    Albert J. Chang

Authors
  1. Wissam Beaino
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  2. Yunjun Guo
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Correspondence to Carolyn J. Anderson.

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Beaino, W., Guo, Y., Chang, A.J. et al. Roles of Atox1 and p53 in the trafficking of copper-64 to tumor cell nuclei: implications for cancer therapy. J Biol Inorg Chem 19, 427–438 (2014). https://doi.org/10.1007/s00775-013-1087-0

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  • DOI: https://doi.org/10.1007/s00775-013-1087-0

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