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Role of mitochondrial oxidative stress in the apoptosis induced by diospyrin diethylether in human breast carcinoma (MCF-7) cells

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Abstract

Mitochondria and associated oxidative stress have been shown to play critical roles in apoptotic death induced by various stress agents. Previously, we reported the antitumor property of diospyrin (D1), a plant-derived bisnaphthoquinonoid, and its diethylether derivative (D7), which was found to cause apoptotic death in human cancer cell lines. The present study aims to explore the relevant mechanism of apoptosis involving generation of cellular reactive oxygen species (ROS) by D7 in human breast carcinoma (MCF-7) cells. It was found that while D7 inhibited the proliferation of tumor cells, the associated apoptosis induced by D7 was prevented by treating the cells with N-acetyl-l-cysteine (NAC), an antioxidant, and cyclosporine A (CsA), an inhibitor of mitochondrial permeability transition (MPT). Experiments using suitable inhibitors also demonstrated that D7 could alter the electron flow in mitochondrial electron transport chain by affecting target(s) between complex I and complex III, and indicated the probable site of D7-induced generation of ROS. These results were further supported by confocal microscopic observation on changes in mitochondrial organization and shape in cells treated with D7. Taken together, the results of our study clearly suggested that the apoptosis induced by D7 would involve alteration of MPT, cardiolipin peroxidation, migration of Bax from cytosol to mitochondria, decreased expression of Bcl-2, and release of cytochrome c, indicating oxidative mechanism at the mitochondrial level in the tumor cells.

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Acknowledgments

This study was supported by the Board of Radiation and Nuclear Sciences, Department of Atomic Energy, Mumbai (Research Grant No. 2002/37/19/BRNS), and the University Grants Commission, New Delhi. Authors would like to acknowledge the technical assistance of Mr. Manjoor Ali, Radiation Biology and Health Sciences Division, BARC, Mumbai in the acquisition of images in Confocal Microscopy. The authors are grateful to Dr. Nandedkar, National Institute for Research in Reproductive Health, Parel, Mumbai, for providing the facility for carrying out the experiments on FACS analysis, and Mr. Akhilesh Chaurasia, SRF, CSIR, BARC for his help in fluorescence microscopy.

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  1. K. P. Mishra

    Present address: Department of Life Sciences, Mumbai University, Kalina, Mumbai, 400 098, India

Authors and Affiliations

  1. Department of Pharmaceutical Technology, Jadavpur University, Kolkata, 700 032, India

    Binod Kumar & Banasri Hazra

  2. Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Mumbai, 400 085, India

    Amit Kumar, B. N. Pandey & K. P. Mishra

  3. United National Institute of Science & Technology, Naini, Allahabad, 211 010, India

    K. P. Mishra

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  1. Binod Kumar
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  5. Banasri Hazra
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Correspondence to B. N. Pandey or Banasri Hazra.

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Kumar, B., Kumar, A., Pandey, B.N. et al. Role of mitochondrial oxidative stress in the apoptosis induced by diospyrin diethylether in human breast carcinoma (MCF-7) cells. Mol Cell Biochem 320, 185–195 (2009). https://doi.org/10.1007/s11010-008-9920-4

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