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Simulating hypoxia-induced acidic environment in cancer cells facilitates mobilization and redox-cycling of genomic copper by daidzein leading to pro-oxidant cell death: implications for the sensitization of resistant hypoxic cancer cells to therapeutic challenges

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Abstract

This study was conducted to investigate the mechanism of action involved in the anti-cancer activity of daidzein and identification of cancer specific micro-environment as therapeutic target of this secondary metabolite derived from soy. Our data indicated that daidzein induces cellular DNA breakage, anti-proliferative effects and apoptosis in a concentration-dependent manner. We demonstrated that such a daidzein-induced anti-cancer action involves a copper-dependant pathway in which endogenous copper is mobilized by daidzein and redox-cycled to generate reactive oxygen species which act as an upstream signal leading to pro-oxidant cell death. Further in the context of hypoxia being a resistant factor against standard therapies and that an effect secondary to hypoxia is the intracellular acidification, we show that the anticancer activity of daidzein is modulated positively in acidic pH but copper-specific chelator is still able to inhibit daidzein activity. Moreover, an experimental setup of hypoxia mimic (cobalt chloride) revealed an enhanced sensitivity of cancer cells to the cytotoxic effects of daidzein which was neutralized in the presence of neocuproine. The findings support a paradigm shift from the conventional antioxidant property of dietary isoflavones to molecules capable of initiating a pro-oxidant signaling mediated by reactive oxygen species. Further, the clinical relevance of such an action mechanism in cancer chemoprevention is also proposed. This study identified endogenous copper as a molecular target and acidic pH as a modulating factor for the therapeutic activity of daidzein against cancer. The evidence presented highlights the potential of dietary agents as adjuvants to standard therapeutic regimens.

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References

  • Adlercreutz CH, Goldin BR, Gorbach SL, Hockerstedt KA, Watanabe S, Hamalainen EK, Markkanen MH, Makela TH, Wahala KT, Adlercreutz T (1995) Soybean phytoestrogen intake and cancer risk. J Nutr 125:757S–770S

    CAS  PubMed  Google Scholar 

  • Ahmad MS, Fazal F, Rahman A, Hadi SM, Parish JH (1992) Activities of flavonoids for the cleavage of DNA in the presence of Cu(II): correlation with generation of active oxygen species. Carcinogenesis 13:605–608

    Article  CAS  Google Scholar 

  • Ahmad A, Farhan AS, Singh S, Hadi SM (2000) DNA breakage by resveratrol and Cu(II): reaction mechanism and bacteriophage inactivation. Cancer Lett 154:29–37

    Article  CAS  PubMed  Google Scholar 

  • Ahmad A, Syed FA, Singh S, Hadi SM (2005) Prooxidant activity of resveratrol in the presence of copper ions: mutagenicity in plasmid DNA. Toxicol Lett 159:1–12

    Article  CAS  PubMed  Google Scholar 

  • Ahmad A, Wang Z, Kong D, Ali R, Ali S, Banerjee S, Sarkar FH (2011) Platelet-derived growth factor-D contributes to aggressiveness of breast cancer cells by up-regulating Notch and NF-kappaB signaling pathways. Breast Cancer Res Treat 126:15–25

    Article  CAS  PubMed  Google Scholar 

  • Ahsan H, Hadi SM (1998) Strand scission in DNA induced by curcumin in the presence of Cu(II). Cancer Lett 124:23–30

    Article  CAS  PubMed  Google Scholar 

  • Apelgot S, Coppey J, Fromentin A, Guille E, Poupon MF, Roussel A (1986) Altered distribution of copper (64Cu) in tumor-bearing mice and rats. Anticancer Res 6:159–164

    CAS  PubMed  Google Scholar 

  • Barnes S, Peterson G, Grubbs C, Setchell K (1994) Potential role of dietary isoflavones in the prevention of cancer. Adv Exp Med Biol 354:135–147

    Article  CAS  PubMed  Google Scholar 

  • Brewer GJ (2005) Anticopper therapy against cancer and diseases of inflammation and fibrosis. Drug Discov Today 10:1103–1109

    Article  CAS  PubMed  Google Scholar 

  • Cao F, Jin TY, Zhou YF (2006) Inhibitory effect of isoflavones on prostate cancer cells and PTEN gene. Biomed Environ Sci 19:35–41

    CAS  PubMed  Google Scholar 

  • Cemeli E, Schmid TE, Anderson D (2004) Modulation by flavonoids of DNA damage induced by estrogen-like compounds. Environ Mol Mutagen 44:420–426

    Article  CAS  PubMed  Google Scholar 

  • Cho J, Kim D, Lee SK, Lee YJ (2005) Cobalt chloride-induced estrogen receptor α down-regulation involves hypoxia-inducible factor-1α in MCF-7 human breast cancer cells. Mol Endocrinol 19(5):1191–1199

    Article  CAS  PubMed  Google Scholar 

  • Choi EJ, Kim GH (2008) Daidzein causes cell cycle arrest at the G1 and G2/M phases in human breast cancer MCF-7 and MDA-MB-453 cells. Phytomedicine 15:683–690

    Article  CAS  PubMed  Google Scholar 

  • Constantinou AI, Lantvit D, Hawthorne M, Xu X, van Breemen RB, Pezzuto JM (2001) Chemopreventive effects of soy protein and purified soy isoflavones on DMBA-induced mammary tumors in female Sprague-Dawley rats. Nutr Cancer 41:75–81

    Article  CAS  PubMed  Google Scholar 

  • Courtois Y, Fromageot P, Guschlbauer W (1968) Protonated polynucleotide structures. 3. An optical rotatory dispersion study of the protonation of DNA. Eur J Biochem 6:493–501

    Article  CAS  PubMed  Google Scholar 

  • Czene S, Tiback M, Harms-Ringdahl M (1997) pH-dependent DNA cleavage in permeabilized human fibroblasts. Biochem J 323(Pt 2):337–341

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Doll R, Peto R (1981) The causes of cancer: quantitative estimates of avoidable risks of cancer in the United States today. J Natl Cancer Inst 66:1191–1308

    CAS  PubMed  Google Scholar 

  • Dong JY, Qin LQ (2011) Soy isoflavones consumption and risk of breast cancer incidence or recurrence: a meta-analysis of prospective studies. Breast Cancer Res Treat 125:315–323

    Article  CAS  PubMed  Google Scholar 

  • Ebara M, Fukuda H, Hatano R, Saisho H, Nagato Y, Suzuki K, Nakajima K, Yukawa M, Kondo F, Nakayama A, Sakurai H (2000) Relationship between copper, zinc and metallothionein in hepatocellular carcinoma and its surrounding liver parenchyma. J Hepatol 33:415–422

    Article  CAS  PubMed  Google Scholar 

  • Fujisawa S, Atsumi T, Ishihara M, Kadoma Y (2004) Cytotoxicity, ROS-generation activity and radical-scavenging activity of curcumin and related compounds. Anticancer Res 24:563–569

    CAS  PubMed  Google Scholar 

  • Goodman VL, Brewer GJ, Merajver SD (2004) Copper deficiency as an anti-cancer strategy. Endocr Relat Cancer 11:255–263

    Article  CAS  PubMed  Google Scholar 

  • Guo JM, Xiao BX, Dai DJ, Liu Q, Ma HH (2004a) Effects of daidzein on estrogen-receptor-positive and negative pancreatic cancer cells in vitro. World J Gastroenterol 10:860–863

    CAS  PubMed  PubMed Central  Google Scholar 

  • Guo JM, Xiao BX, Liu DH, Grant M, Zhang S, Lai YF, Guo YB, Liu Q (2004b) Biphasic effect of daidzein on cell growth of human colon cancer cells. Food Chem Toxicol 42:1641–1646

    Article  CAS  PubMed  Google Scholar 

  • Guo JM, Kang GZ, Xiao BX, Liu DH, Zhang S (2004c) Effect of daidzein on cell growth, cell cycle, and telomerase activity of human cervical cancer in vitro. Int J Gynecol Cancer 14:882–888

    Article  CAS  PubMed  Google Scholar 

  • Gupte A, Mumper RJ (2009) Elevated copper and oxidative stress in cancer cells as a target for cancer treatment. Cancer Treat Rev 35:32–46

    Article  CAS  PubMed  Google Scholar 

  • Hadi SM, Bhat SH, Azmi AS, Hanif S, Shamim U, Ullah MF (2007) Oxidative breakage of cellular DNA by plant polyphenols: a putative mechanism for anticancer properties. Semin Cancer Biol 17:370–376

    Article  CAS  PubMed  Google Scholar 

  • Hadi SM, Ullah MF, Azmi AS, Ahmad A, Shamim U, Zubair H, Khan HY (2010) Resveratrol mobilizes endogenous copper in human peripheral lymphocytes leading to oxidative DNA breakage: a putative mechanism for chemoprevention of cancer. Pharm Res 27:979–988

    Article  CAS  PubMed  Google Scholar 

  • Hoffman DL, Salter JD, Brookes PS (2007) Response of mitochondrial reactive oxygen species generation to steady-state oxygen tension: implications for hypoxic cell signaling. Am J Physiol Heart Circ Physiol 292:H101–H108

    Article  CAS  PubMed  Google Scholar 

  • Hrgovcic M, Tessmer CF, Thomas FB, Ong PS, Gamble JF, Shullenberger CC (1973) Serum copper observations in patients with malignant lymphoma. Cancer 32:1512–1524

    Article  CAS  PubMed  Google Scholar 

  • Ishida I, Kubo H, Suzuki S, Suzuki T, Akashi S, Inoue K, Maeda S, Kikuchi H, Sasaki H, Kondo T (2002) Hypoxia diminishes toll-like receptor 4 expression through reactive oxygen species generated by mitochondria in endothelial cells. J Immunol 169:2069–2075

    Article  CAS  PubMed  Google Scholar 

  • Jin S, Zhang QY, Kang XM, Wang JX, Zhao WH (2010) Daidzein induces MCF-7 breast cancer cell apoptosis via the mitochondrial pathway. Ann Oncol 21:263–268

    Article  CAS  PubMed  Google Scholar 

  • Ju YH, Fultz J, Allred KF, Doerge DR, Helferich WG (2006) Effects of dietary daidzein and its metabolite, equol, at physiological concentrations on the growth of estrogen-dependent human breast cancer (MCF-7) tumors implanted in ovariectomized athymic mice. Carcinogenesis 27:856–863

    Article  CAS  PubMed  Google Scholar 

  • Kagawa TF, Geierstanger BH, Wang AH, Ho PS (1991) Covalent modification of guanine bases in double-stranded DNA. The 1.2-A Z-DNA structure of d(CGCGCG) in the presence of CuCl2. J Biol Chem 266:20175–20184

    CAS  PubMed  Google Scholar 

  • Khan NS, Hadi SM (1998) Structural features of tannic acid important for DNA degradation in the presence of Cu(II). Mutagenesis 13:271–274

    Article  CAS  PubMed  Google Scholar 

  • Kuo HW, Chen SF, Wu CC, Chen DR, Lee JH (2002) Serum and tissue trace elements in patients with breast cancer in Taiwan. Biol Trace Elem Res 89:1–11

    Article  CAS  PubMed  Google Scholar 

  • Liggins J, Bluck LJ, Runswick S, Atkinson C, Coward WA, Bingham SA (2000) Daidzein and genistein content of fruits and nuts. J Nutr Biochem 11:326–331

    Article  CAS  PubMed  Google Scholar 

  • Liggins J, Mulligan A, Runswick S, Bingham SA (2002) Daidzein and genistein content of cereals. Eur J Clin Nutr 56:961–966

    Article  CAS  PubMed  Google Scholar 

  • Lo FH, Mak NK, Leung KN (2007) Studies on the anti-tumor activities of the soy isoflavone daidzein on murine neuroblastoma cells. Biomed Pharmacother 61:591–595

    Article  CAS  PubMed  Google Scholar 

  • Malik A, Azam S, Hadi N, Hadi SM (2003) DNA degradation by water extract of green tea in the presence of copper ions: implications for anticancer properties. Phytother Res 17:358–363

    Article  CAS  PubMed  Google Scholar 

  • Margalioth EJ, Udassin R, Cohen C, Maor J, Anteby SO, Schenker JG (1987) Serum copper level in gynecologic malignancies. Am J Obstet Gynecol 157:93–96

    Article  CAS  PubMed  Google Scholar 

  • Mishra R, Bhadauria S, Murthy PK, Murthy PS (2011) Glycine soya diet synergistically enhances the suppressive effect of tamoxifen and inhibits tamoxifen-promoted hepatocarcinogenesis in 7,12-dimethylbenz[alpha]anthracene-induced rat mammary tumor model. Food Chem Toxicol 49:434–440

    Article  CAS  PubMed  Google Scholar 

  • Moulder JE, Rockwell S (1987) Tumor hypoxia: its impact on cancer therapy. Cancer Metastasis Rev 5:313–341

    Article  CAS  PubMed  Google Scholar 

  • Nemeikaite-Ceniene A, Imbrasaite A, Sergediene E, Cenas N (2005) Quantitative structure-activity relationships in prooxidant cytotoxicity of polyphenols: role of potential of phenoxyl radical/phenol redox couple. Arch Biochem Biophys 441:182–190

    Article  CAS  PubMed  Google Scholar 

  • Nordsmark M, Bentzen SM, Rudat V, Brizel D, Lartigau E, Stadler P, Becker A, Adam M, Molls M, Dunst J, Terris DJ, Overgaard J (2005) Prognostic value of tumor oxygenation in 397 head and neck tumors after primary radiation therapy. An international multi-center study. Radiother Oncol 77:18–24

    Article  PubMed  Google Scholar 

  • Okunieff P, Dunphy EP, Hoeckel M, Terris DJ, Vaupel P (1994) The role of oxygen tension distribution on the radiation response of human breast carcinoma. Adv Exp Med Biol 345:485–492

    Article  CAS  PubMed  Google Scholar 

  • Paddenberg R, Ishaq B, Goldenberg A, Faulhammer P, Rose F, Weissmann N, Braun-Dullaeus RC, Kummer W (2003) Essential role of complex II of the respiratory chain in hypoxia-induced ROS generation in the pulmonary vasculature. Am J Physiol Lung Cell Mol Physiol 284:L710–L719

    Article  CAS  PubMed  Google Scholar 

  • Park OJ, Surh YJ (2004) Chemopreventive potential of epigallocatechin gallate and genistein: evidence from epidemiological and laboratory studies. Toxicol Lett 150:43–56

    Article  CAS  PubMed  Google Scholar 

  • Peng F, Lu X, Janisse J, Muzik O, Shields AF (2006) PET of human prostate cancer xenografts in mice with increased uptake of 64CuCl2. J Nucl Med 47:1649–1652

    CAS  PubMed  Google Scholar 

  • Sakihama Y, Cohen MF, Grace SC, Yamasaki H (2002) Plant phenolic antioxidant and prooxidant activities: phenolics-induced oxidative damage mediated by metals in plants. Toxicology 177:67–80

    Article  CAS  PubMed  Google Scholar 

  • Semczuk B, Pomykalski M (1973) Serum copper level in patients with laryngeal carcinoma. Otolaryngol Pol 27:17–23

    CAS  PubMed  Google Scholar 

  • Semenza GL, Agani F, Iyer N, Jiang BH, Leung S, Wiener C, Yu A (1998) Hypoxia-inducible factor 1: from molecular biology to cardiopulmonary physiology. Chest 114:40S–45S

    Article  CAS  PubMed  Google Scholar 

  • Siegel R, Ma J, Zou Z, Jemal A (2014) Cancer statistics, 2014. CA 64:9–29

    PubMed  Google Scholar 

  • Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175:184–191

    Article  CAS  PubMed  Google Scholar 

  • Singh-Gupta V, Zhang H, Yunker CK, Ahmad Z, Zwier D, Sarkar FH, Hillman GG (2010) Daidzein effect on hormone refractory prostate cancer in vitro and in vivo compared to genistein and soy extract: potentiation of radiotherapy. Pharm Res 27:1115–1127

    Article  CAS  PubMed  Google Scholar 

  • Su SJ, Yeh TM, Lei HY, Chow NH (2006) The potential of soybean foods as a chemoprevention approach for human urinary tract cancer. Clin Cancer Res 6:230–236

    Google Scholar 

  • Sutherland RM, Ausserer WA, Murphy BJ, Laderoute KR (1996) Tumor hypoxia and heterogeneity: challenges and opportunities for the future. Semin Radiat Oncol 6:59–70

    Article  PubMed  Google Scholar 

  • Tani P, Kokkola K (1972) Serum iron, copper, and iron-binding capacity in bronchogenic pulmonary carcinoma. Scand J Respir Dis Suppl 80:121–128

    CAS  PubMed  Google Scholar 

  • Thomasset SC, Berry DP, Garcea G, Marczylo T, Steward WP, Gescher AJ (2007) Dietary polyphenolic phytochemicals–promising cancer chemopreventive agents in humans? A review of their clinical properties. Int J Cancer 120:451–458

    Article  CAS  PubMed  Google Scholar 

  • Ullah MF, Shamim U, Hanif S, Azmi AS, Hadi SM (2009) Cellular DNA breakage by soy isoflavone genistein and its methylated structural analogue biochanin A. Mol Nutr Food Res 53:1376–1385

    Article  CAS  PubMed  Google Scholar 

  • Ullah MF, Ahmad A, Zubair H, Khan HY, Wang Z, Sarkar FH, Hadi SM (2011a) Soy isoflavone genistein induces cell death in breast cancer cells through mobilization of endogenous copper ions and generation of reactive oxygen species. Mol Nutr Food Res 55:553–559

    Article  CAS  PubMed  Google Scholar 

  • Ullah MF, Khan HY, Zubair H, Shamim U, Hadi SM (2011b) The antioxidant ascorbic acid mobilizes nuclear copper leading to a prooxidant breakage of cellular DNA: implications for chemotherapeutic action against cancer. Cancer Chemother Pharmacol 67:103–110

    Article  CAS  PubMed  Google Scholar 

  • Wang GL, Semenza GL (1993) Desferrioxamine induces erythropoietin gene expression and hypoxia-inducible factor 1 DNA-binding activity: implications for models of hypoxia signal transduction. Blood 82:3610–3615

    CAS  PubMed  Google Scholar 

  • Wike-Hooley JL, van den Berg AP, van der Zee J, Reinhold HS (1985) Human tumour pH and its variation. Eur J Cancer Clin Oncol 21:785–791

    Article  CAS  PubMed  Google Scholar 

  • Yoshida D, Ikeda Y, Nakazawa S (1993) Quantitative analysis of copper, zinc and copper/zinc ratio in selected human brain tumors. J Neurooncol 16:109–115

    Article  CAS  PubMed  Google Scholar 

  • Zheng LF, Wei QY, Cai YJ, Fang JG, Zhou B, Yang L, Liu ZL (2006) DNA damage induced by resveratrol and its synthetic analogues in the presence of Cu (II) ions: mechanism and structure-activity relationship. Free Radic Biol Med 41:1807–1816

    Article  CAS  PubMed  Google Scholar 

  • Zuo XL, Chen JM, Zhou X, Li XZ, Mei GY (2006) Levels of selenium, zinc, copper, and antioxidant enzyme activity in patients with leukemia. Biol Trace Elem Res 114:41–53

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry, Faculty of Life Sciences, AMU, Aligarh, 202002, India

    Mohammad F. Ullah, Aamir Ahmad, Husain Y. Khan, Haseeb Zubair & Sheikh M. Hadi

  2. Laboratory of Phytomedicine & Therapeutics, Prince Fahd Research Chair, Department of Medical Laboratory Technology, Faculty of Applied Medical Sciences, University of Tabuk, Tabuk, 71491, Saudi Arabia

    Mohammad F. Ullah & Showket H. Bhat

  3. Department of Pathology, Karmanos Cancer Institute, Wayne State University School of Medicine, Detroit, MI, USA

    Aamir Ahmad & Fazlul H. Sarkar

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  1. Mohammad F. Ullah
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  2. Aamir Ahmad
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  4. Husain Y. Khan
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Correspondence to Mohammad F. Ullah.

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Ullah, M.F., Ahmad, A., Bhat, S.H. et al. Simulating hypoxia-induced acidic environment in cancer cells facilitates mobilization and redox-cycling of genomic copper by daidzein leading to pro-oxidant cell death: implications for the sensitization of resistant hypoxic cancer cells to therapeutic challenges. Biometals 29, 299–310 (2016). https://doi.org/10.1007/s10534-016-9916-6

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