Advertisement

European Journal of Nutrition

, Volume 53, Issue 1, pp 345–350 | Cite as

Daidzein, R-(+)equol and S-(−)equol inhibit the invasion of MDA-MB-231 breast cancer cells potentially via the down-regulation of matrix metalloproteinase-2

  • Pamela J. MageeEmail author
  • Philip Allsopp
  • Adile Samaletdin
  • Ian R. Rowland
Short Communication

Abstract

Purpose

Soy isoflavones may inhibit tumor cell invasion and metastasis via their effects on matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs). The current study investigates the effects of daidzein, R- and S-equol on the invasion of MDA-MB-231 human breast cancer cells and the effects of these compounds on MMP/TIMP expression at the mRNA level.

Methods

The anti-invasive effects of daidzein, R- and S-equol (0, 2.5, 10, 50 μM) on MDA-MB-231 cells were determined using the Matrigel invasion assay following 48-h exposure. Effects on MMP-2, MMP-9, TIMP-1 and TIMP-2 expression were assessed using real-time PCR. Chiral HPLC analysis was used to determine intracellular concentrations of R- and S-equol.

Results

The invasive capacity of MDA-MB-231 cells was significantly reduced (by approximately 50–60 %) following treatment with 50 μM daidzein, R- or S-equol. Anti-invasive effects were also observed with R-equol at 2.5 and 10 μM though overall equipotent effects were induced by all compounds. Inhibition of invasion induced by all three compounds at 50 μM was associated with the down-regulation of MMP-2, while none of the compounds tested significantly affected the expression levels of MMP-9, TIMP-1 or TIMP-2 at this concentration. Following exposure to media containing 50 μM R- or S-equol for 48-h intracellular concentrations of R- and S-equol were 4.38 ± 1.17 and 3.22 ± 0.47 nM, respectively.

Conclusion

Daidzein, R- and S-equol inhibit the invasion of MDA-MB-231 human breast cancer cells in part via the down-regulation of MMP-2 expression, with equipotent effects observed for the parent isoflavone daidzein and the equol enantiomers.

Keywords

Daidzein R-equol S-equol Breast cancer Soy isoflavone 

Notes

Acknowledgments

The authors thank Prof. Herman Adlercreutz for his assistance in facilitating the analysis of intracellular R- and S-equol and Julie McAlinden (Roche Diagnostics Ltd.) for her help with the real-time PCR analysis.

Conflict of interest

I.R. is on the Scientific Advisory Boards of the European Natural Soybean Association, Dean Foods, Alpro Foundation.

References

  1. 1.
    Ferlay J, Shin HR, Bray F, Forman D, Mathers C, Parkin DM (2008) GLOBOCAN 2008, Cancer incidence and mortality worldwide: IARC CancerBase No. 10 [Internet]. Lyon, France: International Agency for Research on Cancer; 2010. Available from: http://globocan.iarc.fr (accessed October 2012)
  2. 2.
    Magee PJ, Rowland IR (2004) Phyto-oestrogens, their mechanism of action: current evidence for a role in breast and prostate cancer. Br J Nutr 91:513–531CrossRefGoogle Scholar
  3. 3.
    Steiner C, Arnould S, Scalbert A, Manach C (2008) Isoflavones and the prevention of breast and prostate cancer: new perspectives opened by nutrigenomics. Br J Nutr 99:ES78–ES108CrossRefGoogle Scholar
  4. 4.
    Yuan J-P, Wang J-H, Liu X (2007) Metabolism of dietary soy isoflavones to equol by human intestinal microflora—implications for health. Mol Nutr Food Res 51:765–781CrossRefGoogle Scholar
  5. 5.
    Setchell KDR, Clerici C, Lephart E, Cole SJ, Heenan C et al (2005) S-equol, a potent ligand for estrogen receptor β, is the exclusive enantiomeric form of the soy isoflavone metabolite produced by human intestinal bacterial flora. Am J Clin Nutr 81:1072–1079Google Scholar
  6. 6.
    Lampe JW, Karr SC, Hutchins AM, Slavin JL (1998) Urinary equol excretion with a soy challenge: influence of habitual diet. Proc Soc Exp Biol Med 217:335–339CrossRefGoogle Scholar
  7. 7.
    Rowland I, Wiseman H, Sanders TAB, Adlercreutz H, Bowey EA (2000) Interindividual variation in metabolism of soy isoflavones and lignans: influence of habitual diet on equol production by the gut microflora. Nutr Cancer 36:27–32CrossRefGoogle Scholar
  8. 8.
    Setchell KDR, Cole SJ (2006) Method of defining equol-producer status and its frequency among vegetarians. J Nutr 136:2188–2193Google Scholar
  9. 9.
    Radisky ES, Radisky DC (2010) Matrix metalloproteinase-induced epithelial-mesenchymal transition in breast cancer. J Mammary Gland Biol Neoplasia 15:201–212CrossRefGoogle Scholar
  10. 10.
    Xu X, Wang Y, Chen Z, Sternlicht M, Hidalgo M, Steffensen B (2005) Matrix metalloproteinase-2 contributes to cancer cell migration on collagen. Cancer Res 65:130–136CrossRefGoogle Scholar
  11. 11.
    Deryugina E, Luo G, Reisfeld R, Bourdon M, Strongin A (1997) Tumor cell invasion through matrigel is regulated by activated matrix metalloproteinase-2. Anticancer Res 17:3201–3210Google Scholar
  12. 12.
    Balduyck M, Zerimech F, Gouyer V, Lemaire R, Hemon B, Grard G, Thiebaut C, Lemaire V, Dacquembronne E, Duhem T, Lebrun A, Dejonghe M, Huet G (2000) Specific expression of matrix metalloproteinases 1, 3, 9 and 13 associated with invasiveness of breast cancer cells in vitro. Clin Exp Metastasis 18:171–178CrossRefGoogle Scholar
  13. 13.
    Azzam H, Arand G, Lippman M, Thompson E (1993) Association of MMP-2 activation potential with metastatic progression in human breast cancer cell lines independent of MMP-2 production. J Natl Cancer Inst 85:1758–1764CrossRefGoogle Scholar
  14. 14.
    Ranogajec I, Jakic-Razumovic J, Puzovic V, Gabrilovac J (2011) Prognostic value of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9) and aminopeptidase N/CD13 in breast cancer patients. Med Oncol May 25 [Epub ahead of print]Google Scholar
  15. 15.
    Leppa S, Saarto T, Vehmanen L, Blomqvist C, Elomaa I (2004) A high serum matrix metalloproteinase-2 level is associated with an adverse prognosis in node-positive breast carcinoma. Clin Cancer Res 10:1057–1063CrossRefGoogle Scholar
  16. 16.
    Rocca GL, Pucci-Minafra I, Marrazzo A, Taormina P, Minafra S (2004) Zymographic detection and clinical correlations of MMP-2 and MMP-9 in breast cancer sera. Br J Cancer 90:1414–1421CrossRefGoogle Scholar
  17. 17.
    Shao Z, Wu J, Shen Z, Barsky S (1998) Genistein exerts multiple suppressive effects on human breast carcinoma cells. Cancer Res 58:4851–4857Google Scholar
  18. 18.
    Magee PJ, McGlynn H, Rowland IR (2004) Differential effects of isoflavones and lignans on invasiveness of MDA-MB-231 breast cancer cells in vitro. Cancer Lett 208:35–41CrossRefGoogle Scholar
  19. 19.
    Magee PJ, Raschke M, Steiner C, Duffin JG, Pool-Zobel BL, Jokela T, Wahala K, Rowland IR (2006) Equol: A comparison of the effects of the racemic compound with that of the purified S-enantiomer on the growth, invasion, and DNA integrity of breast and prostate cells in vitro. Nutr Cancer 54:232–242CrossRefGoogle Scholar
  20. 20.
    Kousidou OC, Mitropoulou TN, Roussidis AE, Kletsas D, Theocharis AD, Karamanos NK (2005) Genistein suppresses the invasive potential of human breast cancer cells through transcriptional regulation of metalloproteinases and their tissue inhibitors. Int J Oncol 26:1101–1109Google Scholar
  21. 21.
    Cailleau R, Young R, Olive M, Reeves W (1974) Breast tumour cell lines from pleural effusions. J Natl Cancer Inst 53:661–674Google Scholar
  22. 22.
    Mossman T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63CrossRefGoogle Scholar
  23. 23.
    Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29(9):e45CrossRefGoogle Scholar
  24. 24.
    Yang Z, Strickland D, Bornstein P (2001) Extracellular matrix metalloproteinase 2 levels are regulated by the low density lipoprotein-related scavenger receptor and thrombospondin 2. J Biol Chem 276:8403–8408CrossRefGoogle Scholar
  25. 25.
    (2011) Anti-invasion effects of R- and S-enantiomers of equol on prostate cancer PC3, DU145 cells. Wei Sheng Yan Jiu 40:423–430 (article in Chinese; no authors listed)Google Scholar
  26. 26.
    Zheng W, Zhang Y, Ma D, Shi Y, Liu C, Wang P (2012) (±) Equol inhibits invasion in prostate cancer DU145 cells possibly via down-regulation of matrix metalloproteinase-9, matrix metalloproteinase-2 and urokinase-type plasminogen activator by antioxidant activity. J Clin Biochem Nutr 51:61–67CrossRefGoogle Scholar
  27. 27.
    Valachovicova T, Slivova V, Bergman H, Shuherk J, Sliva D (2004) Soy isoflavones suppress invasiveness of breast cancer cells by the inhibition of NF-kB/AP-1-dependent and -independent pathways. Int J Oncol 25:1389–1395Google Scholar
  28. 28.
    Brown NM, Belles CA, Lindley SL, Zimmer-Nechemias LD, Zhao X, Witte DP, Kim M-O, Setchell KDR (2010) The chemopreventive action of equol enantiomers in a chemically induced animal model of breast cancer. Carcinogenesis 31:886–893CrossRefGoogle Scholar
  29. 29.
    Maubach J, Depypere HT, Goeman J, Van Der Eycken J, Heyerick A, Bracke ME, Blondeel P, De Keukeleire D (2004) Distribution of soy-derived phytoestrogens in human breast tissue and biological fluids. Obstet Gynecol 103:892–898CrossRefGoogle Scholar
  30. 30.
    Bolca S, Urpi-Sarda M, Blondeel P, Roche N, Vanhaecke L, Possemiers S, Al-Maharik N, Botting N, De Keukeleire D, Bracke M, Heyerick A, Manach C, Depypere H (2010) Disposition of soy isoflavones in normal human breast tissue. Am J Clin Nutr 91:976–984CrossRefGoogle Scholar
  31. 31.
    Setchell KDR, Brown NM, Zhao X, Lindley SL, Heubi JE, King EC, Messina MJ (2011) Soy isoflavone phase II metabolism differs between rodents and humans: implications for the effect on breast cancer risk. Am J Clin Nutr 94:1284–1294CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Pamela J. Magee
    • 1
    Email author
  • Philip Allsopp
    • 1
  • Adile Samaletdin
    • 2
  • Ian R. Rowland
    • 3
  1. 1.School of Biomedical Sciences, Northern Ireland Centre for Food and HealthUniversity of UlsterColeraineNorthern Ireland
  2. 2.Division of Cardiology, Department of MedicineUniversity of HelsinkiHelsinkiFinland
  3. 3.Department of Food and Nutritional SciencesUniversity of ReadingReadingUK

Personalised recommendations