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Critical role for reactive oxygen species in apoptosis induction and cell migration inhibition by diallyl trisulfide, a cancer chemopreventive component of garlic

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Abstract

Diallyl trisulfide (DATS) is a structurally simple but biologically active constituent of processed garlic with in vivo activity against chemically induced as well as oncogene-driven cancer in experimental rodents. This study offers novel insights into the mechanisms underlying anticancer effects of DATS using human breast cancer cells as a model. Exposure of human breast cancer cells (MCF-7 and MDA-MB-231) and a cell line derived from spontaneously developing mammary tumor of a transgenic mouse (BRI-JM04) to DATS resulted in a dose-dependent inhibition of cell viability that was accompanied by apoptosis induction. A non-tumorigenic normal human mammary cell line (MCF-10A) was resistant to growth inhibition and apoptosis induction by DATS. The DATS-induced apoptosis in MDA-MB-231, MCF-7, and BRI-JM04 cells was associated with reactive oxygen species (ROS) production as evidenced by fluorescence microscopy and flow cytometry using a chemical probe (MitoSOX Red). Overexpression of Cu,Zn-superoxide dismutase (Cu,Zn-SOD) as well as Mn-SOD conferred significant protection against DATS-induced ROS production and apoptotic cell death in MDA-MB-231 and MCF-7 cells. Activation of Bak, but not Bax, resulting from DATS treatment was markedly suppressed by overexpression of Mn-SOD. The DATS treatment caused ROS generation, but not activation of Bax or Bak, in MCF-10A cells. Furthermore, the DATS-mediated inhibition of cell migration was partially but significantly attenuated by Cu,Zn-SOD and Mn-SOD overexpression in association with changes in levels of proteins involved in epithelial–mesenchymal transition. The DATS-mediated induction of heme oxygenase-1 was partially attenuated by overexpression of Mn-SOD. These results provide novel mechanistic insights indicating a critical role for ROS in anticancer effects of DATS.

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Abbreviations

DATS:

Diallyl trisulfide

OSCs:

Organosulfides

ROS:

Reactive oxygen species

DAPI:

4′,6-Diamidino-2-phenylindole

DMSO:

Dimethyl sulfoxide

PBS:

Phosphate-buffered saline

SOD:

Superoxide dismutase

HER-2:

Human epidermal growth factor receptor-2

EMT:

Epithelial–mesenchymal transition

JNK:

c-Jun NH2-terminal kinase

HO-1:

Heme oxygenase-1

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Acknowledgments

Research reported in this publication was supported by the National Cancer Institute of the National Institutes of Health under award number R01 CA113363-07 (to SVS). This research project used the Flow Cytometry Facility that was supported in part by a Grant from the National Cancer Institute at the National Institutes of Health under award number P30 CA047904.

Conflict of interest

KC-K, JL, and SVS declare no conflict of interest.

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Authors and Affiliations

  1. Department of Pharmacology & Chemical Biology, University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, 2.32A Hillman Cancer Center Research Pavilion, 5117 Centre Avenue, Pittsburgh, PA, 15213, USA

    Kumar Chandra-Kuntal, Joomin Lee & Shivendra V. Singh

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  1. Kumar Chandra-Kuntal
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  2. Joomin Lee
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  3. Shivendra V. Singh
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Correspondence to Shivendra V. Singh.

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Chandra-Kuntal, K., Lee, J. & Singh, S.V. Critical role for reactive oxygen species in apoptosis induction and cell migration inhibition by diallyl trisulfide, a cancer chemopreventive component of garlic. Breast Cancer Res Treat 138, 69–79 (2013). https://doi.org/10.1007/s10549-013-2440-2

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  • DOI: https://doi.org/10.1007/s10549-013-2440-2

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