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Breast Cancer Research and Treatment

, Volume 138, Issue 1, pp 69–79 | Cite as

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

  • Kumar Chandra-Kuntal
  • Joomin Lee
  • Shivendra V. Singh
Preclinical study

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.

Keywords

Diallyl trisulfide Reactive oxygen species Apoptosis Chemoprevention 

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

Notes

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.

References

  1. 1.
    Challier B, Perarnau JM, Viel JF (1998) Garlic, onion and cereal fibre as protective factors for breast cancer: a French case-control study. Eur J Epidemiol 14:737–747PubMedCrossRefGoogle Scholar
  2. 2.
    Fleischauer AT, Poole C, Arab L (2000) Garlic consumption and cancer prevention: meta-analyses of colorectal and stomach cancers. Am J Clin Nutr 72:1047–1052PubMedGoogle Scholar
  3. 3.
    Block E (1992) The organosulfur chemistry of the genus Allium—implications for the organic chemistry of sulfur. Angew Chem Int Ed Engl 31:1135–1178CrossRefGoogle Scholar
  4. 4.
    Antony ML, Singh SV (2011) Molecular mechanisms and targets of cancer chemoprevention by garlic-derived bioactive compound diallyl trisulfide. Indian J Exp Biol 49:805–816PubMedGoogle Scholar
  5. 5.
    Xiao D, Choi S, Johnson DE et al (2004) Diallyl trisulfide-induced apoptosis in human prostate cancer cells involves c-Jun N-terminal kinase and extracellular-signal regulated kinase-mediated phosphorylation of Bcl-2. Oncogene 23:5594–5606PubMedCrossRefGoogle Scholar
  6. 6.
    Na HK, Kim EH, Choi MA, Park JM, Kim DH, Surh YJ (2012) Diallyl trisulfide induces apoptosis in human breast cancer cells through ROS-mediated activation of JNK and AP-1. Biochem Pharmacol 84:1241–1250PubMedCrossRefGoogle Scholar
  7. 7.
    Singh SV, Mohan RR, Agarwal R et al (1996) Novel anti-carcinogenic activity of an organosulfide from garlic: inhibition of H-RAS oncogene transformed tumor growth in vivo by diallyl disulfide is associated with inhibition of p21-H-ras processing. Biochem Biophys Res Commun 225:660–665PubMedCrossRefGoogle Scholar
  8. 8.
    Sparnins VL, Barany G, Wattenberg LW (1988) Effects of organosulfur compounds from garlic and onions on benzo[a]pyrene-induced neoplasia and glutathione S-transferase activity in the mouse. Carcinogenesis 9:131–134PubMedCrossRefGoogle Scholar
  9. 9.
    Xiao D, Lew KL, Kim YA et al (2006) Diallyl trisulfide suppresses growth of PC-3 human prostate cancer xenograft in vivo in association with Bax and Bak induction. Clin Cancer Res 12:6836–6843PubMedCrossRefGoogle Scholar
  10. 10.
    Wu PP, Liu KC, Huang WW et al (2011) Diallyl trisulfide (DATS) inhibits mouse colon tumor in mouse CT-26 cells allograft model in vivo. Phytomedicine 18:672–676PubMedCrossRefGoogle Scholar
  11. 11.
    Singh SV, Powolny AA, Stan SD et al (2008) Garlic constituent diallyl trisulfide prevents development of poorly-differentiated prostate cancer and pulmonary metastasis multiplicity in TRAMP mice. Cancer Res 68:9503–9511PubMedCrossRefGoogle Scholar
  12. 12.
    Shrotriya S, Kundu JK, Na HK, Surh YJ (2010) Diallyl trisulfide inhibits phorbol ester-induced tumor promotion, activation of AP-1, and expression of COX-2 in mouse skin by blocking JNK and Akt signaling. Cancer Res 70:1932–1940PubMedCrossRefGoogle Scholar
  13. 13.
    Xiao D, Li M, Herman-Antosiewicz A et al (2006) Diallyl trisulfide inhibits angiogenic features of human umbilical vein endothelial cells by causing Akt inactivation and down-regulation of VEGF and VEGF-R2. Nutr Cancer 55:94–107PubMedCrossRefGoogle Scholar
  14. 14.
    Xiao D, Herman-Antosiewicz A, Antosiewicz J et al (2005) Diallyl trisulfide-induced G2-M phase cell cycle arrest in human prostate cancer cells is caused by reactive oxygen species-dependent destruction and hyperphosphorylation of Cdc25C. Oncogene 24:6256–6268PubMedCrossRefGoogle Scholar
  15. 15.
    Herman-Antosiewicz A, Singh SV (2005) Checkpoint kinase 1 regulates diallyl trisulfide-induced mitotic arrest in human prostate cancer cells. J Biol Chem 280:28519–28528PubMedCrossRefGoogle Scholar
  16. 16.
    Herman-Antosiewicz A, Stan SD, Hahm ER, Xiao D, Singh SV (2007) Activation of a novel ataxia-telangiectasia mutated and Rad3 related/checkpoint kinase 1-dependent prometaphase checkpoint in cancer cells by diallyl trisulfide, a promising cancer chemopreventive constituent of processed garlic. Mol Cancer Ther 6:1249–1261PubMedCrossRefGoogle Scholar
  17. 17.
    Kim YA, Xiao D, Xiao H et al (2007) Mitochondria-mediated apoptosis by diallyl trisulfide in human prostate cancer cells is associated with generation of reactive oxygen species and regulated by Bax/Bak. Mol Cancer Ther 6:1599–1609PubMedCrossRefGoogle Scholar
  18. 18.
    Xiao D, Singh SV (2006) Diallyl trisulfide, a constituent of processed garlic, inactivates Akt to trigger mitochondrial translocation of BAD and caspase-mediated apoptosis in human prostate cancer cells. Carcinogenesis 27:533–540PubMedCrossRefGoogle Scholar
  19. 19.
    Xiao D, Vogel V, Singh SV (2006) Benzyl isothiocyanate-induced apoptosis in human breast cancer cells is initiated by reactive oxygen species and regulated by Bax and Bak. Mol Cancer Ther 5:2931–2945PubMedCrossRefGoogle Scholar
  20. 20.
    Hahm ER, Moura MB, Kelley EE, van Houten B, Shiva S, Singh SV (2011) Withaferin A-induced apoptosis in human breast cancer cells is mediated by reactive oxygen species. PLoS ONE 6:e23354PubMedCrossRefGoogle Scholar
  21. 21.
    Kim SH, Sehrawat A, Sakao K, Hahm ER, Singh SV (2011) Notch activation by phenethyl isothiocyanate attenuates its inhibitory effect on prostate cancer cell migration. PLoS ONE 6:e26615PubMedCrossRefGoogle Scholar
  22. 22.
    Xiao D, Srivastava SK, Lew KL et al (2003) Allyl isothiocyanate, a constituent of cruciferous vegetables, inhibits proliferation of human prostate cancer cells by causing G2/M arrest and inducing apoptosis. Carcinogenesis 24:891–897PubMedCrossRefGoogle Scholar
  23. 23.
    Powolny AA, Bommareddy A, Hahm ER et al (2011) Chemopreventative potential of the cruciferous vegetable constituent phenethyl isothiocyanate in a mouse model of prostate cancer. J Natl Cancer Inst 103:571–584PubMedCrossRefGoogle Scholar
  24. 24.
    Singh SV, Srivastava SK, Choi S et al (2005) Sulforaphane-induced cell death in human prostate cancer cells is initiated by reactive oxygen species. J Biol Chem 280:19911–19924PubMedCrossRefGoogle Scholar
  25. 25.
    Xiao D, Powolny AA, Moura MB et al (2010) Phenethyl isothiocyanate inhibits oxidative phosphorylation to trigger reactive oxygen species-mediated death of human prostate cancer cells. J Biol Chem 285:26558–26569PubMedCrossRefGoogle Scholar
  26. 26.
    Xiao D, Powolny AA, Singh SV (2008) Benzyl isothiocyanate targets mitochondrial respiratory chain to trigger reactive oxygen species-dependent apoptosis in human breast cancer cells. J Biol Chem 283:30151–30163PubMedCrossRefGoogle Scholar
  27. 27.
    Buccellato LJ, Tso M, Akinci OI, Chandel NS, Budinger GR (2004) Reactive oxygen species are required for hyperoxia-induced Bax activation and cell death in alveolar epithelial cells. J Biol Chem 279:6753–6760PubMedCrossRefGoogle Scholar
  28. 28.
    Friedl P, Wolf K (2003) Tumour-cell invasion and migration: diversity and escape mechanism. Nat Rev Cancer 3:362–374PubMedCrossRefGoogle Scholar
  29. 29.
    Tomaskovic-Crook E, Thompson EW, Thiery JP (2009) Epithelial to mesenchymal transition and breast cancer. Breast Cancer Res 11:213PubMedCrossRefGoogle Scholar
  30. 30.
    Werts ED, Gould MN (1986) Relationships between cellular superoxide dismutase and susceptibility to chemically induced cancer in the rat mammary gland. Carcinogenesis 7:1197–1201PubMedCrossRefGoogle Scholar
  31. 31.
    Li JJ, Oberley LW, St Clair DK, Ridnour LA, Oberley TD (1995) Phenotypic changes induced in human breast cancer cells by overexpression of manganese-containing superoxide dismutase. Oncogene 10:1989–2000PubMedGoogle Scholar
  32. 32.
    Tsai SM, Hou MF, Wu SH et al (2011) Expression of manganese superoxide dismutase in patients with breast cancer. Kaohsiung J Med Sci 27:167–172PubMedCrossRefGoogle Scholar
  33. 33.
    Chen C, Pung D, Leong V et al (2004) Induction of detoxifying enzymes by garlic organosulfur compounds through transcription factor Nrf2: effect of chemical structure and stress signals. Free Radic Biol Med 37:1578–1590PubMedCrossRefGoogle Scholar
  34. 34.
    Li H, Li HQ, Wang Y et al (2004) An intervention study to prevent gastric cancer by micro-selenium and large dose of allitridum. Chin Med J 117:1155–1160PubMedGoogle Scholar
  35. 35.
    Sun X, Guo T, He J et al (2006) Simultaneous determination of diallyl trisulfide and diallyl disulfide in rat blood by gas chromatography with electron-capture detection. Pharmazie 61:985–988PubMedGoogle Scholar
  36. 36.
    Antosiewicz J, Herman-Antosiewicz A, Marynowski SW, Singh SV (2006) c-Jun NH2-terminal kinase signaling axis regulates diallyl trisulfide-induced generation of reactive oxygen species and cell cycle arrest in human prostate cancer cells. Cancer Res 66:5379–5386PubMedCrossRefGoogle Scholar
  37. 37.
    Borkowska A, Sielicka-Dudzin A, Herman-Antosiewicz A et al (2012) Diallyl trisulfide-induced prostate cancer cell death is associated with Akt/PKB dephosphorylation mediated by P-p66shc. Eur J Nutr 51:817–825PubMedCrossRefGoogle Scholar
  38. 38.
    Sielicka-Dudzin A, Borkowska A, Herman-Antosiewicz A et al (2012) Impact of JNK1, JNK2, and ligase Itch on reactive oxygen species formation and survival of prostate cancer cells treated with diallyl trisulfide. Eur J Nutr 51:573–581PubMedCrossRefGoogle Scholar
  39. 39.
    Hurd TR, DeGennaro M, Lehmann R (2012) Redox regulation of cell migration and adhesion. Trends Cell Biol 22:107–115PubMedCrossRefGoogle Scholar
  40. 40.
    Tochhawng L, Deng S, Pervaiz S, Yap CT (2012) Redox regulation of cancer cell migration and invasion. Mitochondrion (in press), doi:  10.1016/j.mito.2012.08.002
  41. 41.
    Waris G, Ahsan H (2006) Reactive oxygen species: role in the development of cancer and various chronic conditions. J Carcinog 5:14PubMedCrossRefGoogle Scholar
  42. 42.
    Wu WS, Wu JR, Hu CT (2008) Signal cross talks for sustained MAPK activation and cell migration: the potential role of reactive oxygen species. Cancer Metastasis Rev 27:303–314PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2013

Authors and Affiliations

  • Kumar Chandra-Kuntal
    • 1
  • Joomin Lee
    • 1
  • Shivendra V. Singh
    • 1
  1. 1.Department of Pharmacology & Chemical BiologyUniversity of Pittsburgh Cancer Institute, University of Pittsburgh School of MedicinePittsburghUSA

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