Cell Survival, Apoptosis and AMPK-COX-2 Signaling Pathway of Mammary Tumor Cells after Genistein Treatment Combined with Estrogen

  • Lee, Yun-Kyoung ;
  • Hwang, Jin-Taek ;
  • Kim, Young-Min ;
  • Park, Ock-Jin
  • Published : 2007.12.31


Genistein is an active component of legumes and other related food shown to be associated with prevention of degenerative diseases such as cancer through inducing signaling pathways. Treatment of genistein resulted in the induction of apoptosis in the cultured cancer cells. This induction of apoptosis was demonstrated by the Tunel assay in these cells. Unveiling the potential of genistein in cytotoxicity via apoptosis when it is treated with estrogen can predict the therapeutic capability of genistein in breast cancers in the presence of endogenous estrogen. We have found that apoptosis induced by genistein treatment in the presence of estrogen is agonistic or antagonistic depending on the concentrations and treatment periods applied in MCF-7 breast cancer cells. For the suppression of cell survival, 24 hr of treatment was required to induce a synergistic agonistic response between estrogen and genistein at low concentrations of genistein. After this period, the agonistic pattern of genistein to estrogen disappeared. The decrement of COX-2 expression in MCF-7 cells treated with genistein was accompanied with the activation of AMPK only at a high concentration of genistein. This association between AMPK activation and down-regulation of COX-2 by genistein was dampened in the presence of estrogen. It was also demonstrated that genistein and estrogen regulate cell survival and apoptosis by modulating p53 and caspase-3 in the opposite direction. These results suggest that genistein has the potential to control breast cancer development, and co-treatment with estrogen can cause agonistic or antagonistic action on breast cancer cell control.


genistein;genistein cancer prevention in presence of estrogen;apoptosis;AMPK-COX-2 pathway;apoptotic proteins;MCF-7 breast cancer cells


  1. Brzozowski AM, Pike CW, Dauter Z, Hubbard RE, Bonn T, Engstrom O, Ohman L, Greene GL, Gustafsson J, Carlquist M. 1997. Molecular basis of agonism and antagonism in the oestrogen receptor. Nature 389: 753-758
  2. Ziegler RG, Hoover RN, Pike MC, Hildesheim A, Nomura AM, West DW, Wu-Williams AH, Kolonel LN, Horn- Ross PL, Rosenthal JF, Hyer MB. 1993. Migration patterns and breast cancer risk in Asian-American women. J Nat Cancer Inst 85: 1819-1827
  3. Zheng W, Dai Q, Custer LJ, Shu XO, Wen WQ, Jin F, Franke AA. 1999. Urinary excretion of isoflavonoids and the risk of breast cancer. Cancer Epidemiol Biomark Prev 8: 35-40
  4. Martin PM, Horwitz KB, Ryan DS, McGuire WL. 1978. Phytoestrogen interaction with estrogen receptors in human breast cancer cells. Endocrinology 103: 1860-1867
  5. Hsieh CY, Santell RC, Haslam SZ, Helferich WG. 1998. Estrogenic effects of genistein on the growth of estrogen receptor-positive human breast cancer (MCF-7) cells in vitro and in vivo. Cancer Res 58: 3833-3838
  6. Ju YH, Allred KF, Allred CD, Helferich WG. 2006. Genistein stimulates growth of human breast cancer cells in a novel, postmenopausal animal model, with low plasma estradiol concentrations. Carcinogenesis 27: 1292- 1299
  7. Zava DT, Duwe G. 1997. Estrogenic and antiproliferative properties of genistein and other flavonoids in human breast cancer cells in vitro. Nutr Cancer 27: 31-40
  8. Mai Z, Blackburn GL, Zhou JR. 2007. Soy phytochemicals synergistically enhance the preventive effect of tamoxifen on the growth of estrogen-dependent human breast carcinoma in mice. Carcinogenesis 28: 1217-1223
  9. Willard ST, Frawley LS. 1998. Phytoestrogens have agonistic and combinatorial effects on estrogen-responsive gene expression in MCF-7 human breast cancer cells. Endocrine 8: 117-121
  10. Messina M, McCaskill-Stevens W, Lampe JW. 2006. Addressing the soy and breast cancer relationship: review, commentary, and workshop proceedings. J Natl Cancer Inst 98: 1275-1284
  11. Cappelletti V, Fioravanti L, Miodini P, Di Fronzo G. 2000. Genistein blocks breast cancer cells in the G2M phase of the cell cycle. J Cell Biochem 79: 594-600<594::AID-JCB80>3.0.CO;2-4
  12. Seo HS, DeNardo DG, Jacquot Y, Laios I, Vidal DS, Zambrana CR, Leclercq G, Brown PH. 2006. Stimulatory effect of genistein and apigenin on the growth of breast cancer cells correlates with their ability to activate ER alpha. Breast Cancer Res Treat 99: 121-134
  13. Schmidt S, Michna H, Diel P. 2005. Combinatory effects of phytoestrogens and 17$\beta$-estradiol on proliferation and apoptosis in MCF-7 breast cancer cells. J Steroid Biochem Mol Biol 94: 445-449
  14. Nenci I, Marchetti E, Querzoli P. 1988. Commentary on human mammary preneoplasia. The estrogen receptor-promotion hypothesis. J Steriod Biochem 30: 105-106
  15. Perillo B, Sasso A, Abbondanza C, Palumbo G. 2000. 17 $\beta$-estradiol inhibits apoptosis in MCF-7 cells, including bcl-2 expression via two estrogen-responsive elements present in the coding sequence. Mol Cell Biol 20: 2890- 2901
  16. Liu RH. 2004. Potential synergy of phytochemicals in cancer prevention: mechanism of action. J Nutr 134: 3479S- 3485S
  17. Duthie SJ. 2007. Berry phytochemicals, genomic stability and cancer: evidence for chemoprotection at several stages in the carcinogenic process. Mol Nutr Food Res 51: 665-674
  18. Cline JM, Hughes CL Jr. 1998. Phytochemicals for the prevention of breast and endometrial cancer. Cancer Treat Res 94: 107-134
  19. Hwang JT, Ha J, Park IJ, Lee SK, Baik HW, Kim YM, Park OJ. 2007. Apoptotic effect of EGCG in HT-29 colon cancer cells via AMPK signal pathway. Cancer Lett 247: 115-121
  20. Hwang JT, Kwak DW, Lin SK, Kim HM, Kim YM, Park OJ. 2007. Resveratrol induces apoptosis in chemoresistant cancer cells via modulation of AMPK signaling pathway. Ann N Y Acad Sci 1095: 441-448
  21. Lau TY, Leung LK. 2006. Soya isoflavones suppress phorbol 12-myristate 13-acetate-induced COX-2 expression in MCF-7 cells. Br J Nutr 96: 169-176
  22. Hwang JT, Ha J, Park OJ. 2005. Combination of 5-fluorouracil and genistein induces apoptosis synergistically in chemo-resistant cancer cells through the modulation of AMPK and COX-2 signaling pathways. Biochem Biophys Res Commun 332: 433-440
  23. Xu J, Loo G. 2001. Different effects of genistein on molecular markers related to apoptosis in two phenotypically dissimilar breast cancer cell lines. J Cell Biochem 82: 78-88
  24. Yang S, Zhou Q, Yang X. 2007. Caspase-3 status is a determinant of the differential responses to genistein between MDA-MB-231 and MCF-7 breast cancer cells. Biochim Biophys Acta 1773: 903-911
  25. Russo J, Balogh GA, Heulings R, Mailo DA, Moral R, Russo PA, Sheriff F, Vanegas J, Russo IH. 2006. Molecular basis of pregnancy-induced breast cancer protection. Eur J Cancer Prev 15: 306-342
  26. Houssami N, Cuzick J, Dixon JM. 2006. The prevention, detection, and manage-ment of breast cancer. Med J Aust 184: 230-234
  27. Sarkar FH, Li Y. 2004. The role of isoflavones in cancer chemoprevention. Front Biosci 9: 2714-2724
  28. Kaklamani VG, Pasche B. 2004. Role of TGF-beta in cancer and the potential for therapy and prevention. Expert Rev Anticancer Ther 4: 649-661
  29. Shen Q, Brown PH. 2003. Novel agents for the prevention of breast cancer: targeting transcription factors and signal transduction pathways. J Mammary Gland Biol Neoplasia 8: 45-73
  30. Hwang JT, Kim YM, Surh YJ, BaikHW, Lee SK, Ha J, Park OJ. 2006. Selenium regulates cyclooxygenase-2 and extracellular signal-regulated kinase signaling pathways by activating AMP-activated protein kinase in colon cancer cells. Cancer Research 66: 10057-10063