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Bacterial cupredoxin azurin as an inducer of apoptosis and regression in human breast cancer

Oncogene volume 23, pages 2367–2378 (2004)Cite this article

Abstract

Azurin, a copper-containing redox protein released by the pathogenic bacterium Pseudomonas aeruginosa, is highly cytotoxic to the human breast cancer cell line MCF-7, but is less cytotoxic toward p53-negative (MDA-MB-157) or nonfunctional p53 cell lines like MDD2 and MDA-MB-231. The purpose of this study was to investigate the underlying mechanism of the action of bacterial cupredoxin azurin in the regression of breast cancer and its potential chemotherapeutic efficacy. Azurin enters into the cytosol of MCF-7 cells and travels to the nucleus, enhancing the intracellular levels of p53 and Bax, thereby triggering the release of mitochondrial cytochrome c into the cytosol. This process activates the caspase cascade (including caspase-9 and caspase-7), thereby initiating the apoptotic process. Our results indicate that azurin-induced cell death stimuli are amplified in the presence of p53. In vivo injection of azurin in immunodeficient mice harboring xenografted human breast cancer cells in the mammary fat pad leads to statistically significant regression (85%, P=0.0179, Kruskal–Wallis Test) of the tumor. In conclusion, azurin blocks breast cancer cell proliferation and induces apoptosis through the mitochondrial pathway both in vitro and in vivo, thereby suggesting a potential chemotherapeutic application of this bacterial cupredoxin for the treatment of breast cancer.

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Acknowledgements

We thank Mr Albert Green for helping with the animal studies, Ms Laura Bratescu for maintaining all the cell cultures, and Ms Anne Shilkaitis for helping with immunocytochemical staining of the tumors. Thanks are also due to Drs Thomas Shenk and R Baskaran for genetic constructs, and to Drs N Mahile and CJ Froelich for the cell lines used in this work. The editorial help and supervision of Mr Scott Kennedy in manuscript preparation is acknowledged. This research was supported by US National Institutes of Health Grants: PHS ES04050-17, CA09432, CA96517, CA95712, DOD Grant DAMD17-99-1-9223, NSF Grant DMS-0204532, and a gift from the Raymond Cole Foundation.

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Authors and Affiliations

  1. Department of Surgical Oncology, University of Illinois at Chicago, 840 South Wood Street, Chicago, M/C 820, 60612, IL, USA

    Vasu Punj, Djenann Saint-Dic, Elizabeth A Cunningham, Jewell Graves, Andreas I Constantinou, Konstantin Christov, Bethany White & Tapas K Das Gupta

  2. Department of Microbiology and Immunology, University of Illinois at Chicago, 901 S. Wolcott Ave., M/C 790, Chicago, 60612-7344, IL, USA

    Vasu Punj, Suchita Bhattacharyya, Tohru Yamada & Ananda M Chakrabarty

  3. Department of Surgery, University of Illinois at Chicago, 840 S. Wood St., M/C 958, Chicago, 60612, IL, USA

    Chenthamarakshan Vasu

  4. Department of Mathematics, Statistics, and Computer Sciences, 851 S. Morgan, MC 249, University of Illinois at Chicago, Chicago, 60607, IL, USA

    Gang Li & Dibyen Majumdar

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  1. Vasu Punj
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Correspondence to Tapas K Das Gupta.

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Punj, V., Bhattacharyya, S., Saint-Dic, D. et al. Bacterial cupredoxin azurin as an inducer of apoptosis and regression in human breast cancer. Oncogene 23, 2367–2378 (2004). https://doi.org/10.1038/sj.onc.1207376

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