Quercetin greatly improved therapeutic index of doxorubicin against 4T1 breast cancer by its opposing effects on HIF-1α in tumor and normal cells

  • Gangjun Du
  • Haihong Lin
  • Mei Wang
  • Shuo Zhang
  • Xianchuang Wu
  • Linlin Lu
  • Liyan Ji
  • Lijuan Yu
Original Article

Abstract

Purpose

The anthracycline antibiotic doxorubicin (DOX) has been used successfully for treating various types of cancers. However, the therapeutic efficacy of DOX was greatly restricted by its cumulative dose-related cardiotoxicity and common side effects such as bone marrow and immune suppression. Quercetin had better cardioprotective and hepatoprotective activities. The present study was to observe whether quercetin could improve therapeutic index of DOX and explore its mechanisms.

Methods

Effects of quercetin on doxorubicin (DOX)-induced cytotoxicity were investigated in 4T1 cells and murine spleen cells by methylthiazoletetrazolium assay, flow cytometry and single cell gel electrophoresis. Influences of quercetin on therapeutic efficacy and systemic toxicity of DOX were evaluated in BALB/c mice with 4T1 breast cancer. Hypoxia-inducible factor-1 alpha (HIF-1α) in tumor and normal cells was examined to explore mechanisms of quercetin by Western blot and enzyme-linked immunosorbent assay.

Results

In vitro, quercetin at dose less than 100 μM had only slight effects on cell viability and DOX-induced cytotoxicity in 4T1 cells under normoxia, but it could reverse 4T1 cell resistance to DOX under hypoxia and protect spleen cells against DOX-induced cytotoxicity. In vivo, quercetin suppressed tumor growth and prolonged survival in BALB/c mice bearing 4T1 breast cancer. Importantly, quercetin enhanced therapeutic efficacy of DOX and simultaneously reduced DOX-induced toxic side effects. Further study showed that quercetin suppressed intratumoral HIF-1α in a hypoxia-dependent way but increased its accumulation in normal cells. HIF-1α siRNA abolished effects of quercetin on both tumor and normal cells.

Conclusions

These results suggested that quercetin could improve therapeutic index of DOX by its opposing effects on HIF-1α in tumor and normal cells, and was a promising candidate as anticancer agents.

Keywords

Quercetin Doxorubicin Therapeutic index HIF-1α Breast cancer 

References

  1. 1.
    Jemal A, Siegel R, Ward E, Hao Y, Xu J, Murray T, Thun MJ (2008) Cancer statistics, 2008. CA Cancer J Clin 58:71–96CrossRefPubMedGoogle Scholar
  2. 2.
    Gonzalez-Angulo AM, Morales-Vasquez F, Hortobagyi GN (2007) Overview of resistance to systemic therapy in patients with breast cancer. Adv Exp Med Biol 608:1–22PubMedGoogle Scholar
  3. 3.
    Morse R, Rodgers J, Verrill M, Kendell K (2003) Neuropsychological functioning following systemic treatment in women treated for breast cancer: a review. Eur J Cancer 39:2288–2297CrossRefPubMedGoogle Scholar
  4. 4.
    Gianni L (2009) Anthracyclines and early breast cancer: the end of an era? J Clin Oncol 27:1155–1157CrossRefPubMedGoogle Scholar
  5. 5.
    Gianni L, Salvatorelli E, Minotti G (2007) Anthracycline cardiotoxicity in breast cancer patients: synergism with trastuzumab and taxanes. Cardiovasc Toxicol 7(2):67–71CrossRefPubMedGoogle Scholar
  6. 6.
    Benavente-García O, Castillo J, Alcaraz M, Vicente V, Del Río JA, Ortuño A (2007) Beneficial action of citrus flavonoids on multiple cancer-related biological pathways. Curr Cancer Drug Targets 7:795–809CrossRefPubMedGoogle Scholar
  7. 7.
    Formica JV, Regelson W (1995) Review of the biology of quercetin and related bioflavonoids. Food Chem Toxicol 33:1061–1080CrossRefPubMedGoogle Scholar
  8. 8.
    Hirpara KV, Aggarwal P, Mukherjee AJ, Joshi N, Burman AC (2009) Quercetin and its derivatives: synthesis, pharmacological uses with special emphasis on anti-tumor properties and prodrug with enhanced bio-availability. Anticancer Agents Med Chem 9:138–161PubMedGoogle Scholar
  9. 9.
    Mulholland PJ, Ferry DR, Anderson D, Hussain SA, Young AM, Cook JE, Hodgkin E, Seymour LW, Kerr DJ (2001) Pre-clinical and clinical study of QC12, a water-soluble, pro-drug of quercetin. Ann Oncol 12:245–248CrossRefPubMedGoogle Scholar
  10. 10.
    Damianaki A, Bakogeorgou E, Kampa M, Notas G, Hatzoglou A, Panagiotou S, Gemetzi C, Kouroumalis E, Martin PM, Castanas E (2000) Potent inhibitory action of red wine polyphenols on human breast cancer cells. J Cell Biochem 78:429–441CrossRefPubMedGoogle Scholar
  11. 11.
    Conklin CM, Bechberger JF, MacFabe D, Guthrie N, Kurowska EM, Naus CC (2007) Genistein and quercetin increase connexin43 and suppress growth of breast cancer cells. Carcinogenesis 28:93–100CrossRefPubMedGoogle Scholar
  12. 12.
    Hakimuddin F, Paliyath G, Meckling K (2004) Selective cytotoxicity of a red grape wine flavonoid fraction against MCF-7 cells. Breast Cancer Res Treat 85:65–79CrossRefPubMedGoogle Scholar
  13. 13.
    Verma N, Behera BC, Makhija U (2008) Antioxidant and hepatoprotective activity of a lichen Usnea ghattensis in vitro. Appl Biochem Biotechnol 151:167–181CrossRefPubMedGoogle Scholar
  14. 14.
    Mojzisová G, Mirossay L, Kucerová D, Kyselovic J, Mirossay A, Mojzis J (2006) Protective effect of selected flavonoids on in vitro daunorubicin-induced cardiotoxicity. Phytother Res 20:110–114CrossRefPubMedGoogle Scholar
  15. 15.
    Duthie SJ, Collins AR, Duthie GG, Dobson VL (1997) Quercetin and myricetin protect against hydrogen peroxide-induced DNA damage (strand breaks and oxidised pyrimidines) in human lymphocytes. Mutat Res 393:223–231PubMedGoogle Scholar
  16. 16.
    Granado-Serrano AB, Martín MA, Bravo L, Goya L, Ramos S (2006) Quercetin induces apoptosis via caspase activation, regulation of Bcl-2, and inhibition of PI-3-kinase/Akt and ERK pathways in a human hepatoma cell line (HepG2). J Nutr 136:2715–2721PubMedGoogle Scholar
  17. 17.
    Tang N, Du GJ, Wang N, Liu CC, Hang HY, Liang W (2007) Improving penetration in tumors with nanoassemblies of phospholipids and doxorubicin. J Natl Cancer Inst 99:1004–1015CrossRefPubMedGoogle Scholar
  18. 18.
    Collins AR (2004) The comet assay for DNA damage and repair: principles, applications, and limitations. Mol Biotechnol 26:249–261CrossRefPubMedGoogle Scholar
  19. 19.
    Choi HJ, Eun JS, Kim BG, Kim SY, Jeon H, Soh Y (2006) Vitexin, an HIF-1α inhibitor, has anti-metastatic potential in PC12 cells. Mol Cells 22:291–299PubMedGoogle Scholar
  20. 20.
    Early Breast Cancer Trialists’ Collaborative Group (2005) Effects of chemotherapy and hormonal therapy for early breast cancer on recurrence and 15-year survival: an overview of the randomised trials. Lancet 365:1687–1717CrossRefGoogle Scholar
  21. 21.
    Lee DH, Lee YJ (2008) Quercetin suppresses hypoxia-induced accumulation of hypoxia-inducible factor-1alpha (HIF-1alpha) through inhibiting protein synthesis. J Cell Biochem 105:546–553CrossRefPubMedGoogle Scholar
  22. 22.
    Jeon H, Kim H, Choi D, Kim D, Park SY, Kim YJ, Kim YM, Jung Y (2007) Quercetin activates an angiogenic pathway, hypoxia inducible factor (HIF)-1-vascular endothelial growth factor, by inhibiting HIF-prolyl hydroxylase: a structural analysis of quercetin for inhibiting HIF prolyl hydroxylase. Mol Pharmacol 71:1676–1684CrossRefPubMedGoogle Scholar
  23. 23.
    Triantafyllou A, Liakos P, Tsakalof A, Chachami G, Paraskeva E, Molyvdas PA, Georgatsou E, Simos G, Bonanou S (2007) The flavonoid quercetin induces hypoxia-inducible factor-1alpha (HIF-1alpha) and inhibits cell proliferation by depleting intracellular iron. Free Radic Res 41:342–356CrossRefPubMedGoogle Scholar
  24. 24.
    Middleton E Jr, Kandaswami C, Theoharides TC (2000) The effects of plant flavonoids on mammalian cells: implications for inflammation, heart disease, and cancer. Pharmacol Rev 52:673–751PubMedGoogle Scholar
  25. 25.
    Ruiz MJ, Fernández M, Picó Y, Mañes J, Asensi M, Carda C, Asensio G, Estrela JM (2009) Dietary administration of high doses of pterostilbene and quercetin to mice is not toxic. J Agric Food Chem 57:3180–3186CrossRefPubMedGoogle Scholar
  26. 26.
    Harwood M, Danielewska-Nikiel B, Borzelleca JF, Flamm GW, Williams GM, Lines TC (2007) A critical review of the data related to the safety of quercetin and lack of evidence of in vivo toxicity, including lack of genotoxic/carcinogenic properties. Food Chem Toxicol 45:2179–2205CrossRefPubMedGoogle Scholar
  27. 27.
    Utesch D, Feige K, Dasenbrock J, Broschard TH, Harwood M, Danielewska-Nikiel B, Lines TC (2008) Evaluation of the potential in vivo genotoxicity of quercetin. Mutat Res 654:38–44PubMedGoogle Scholar
  28. 28.
    Scambia G, Ranelletti FO, Panici PB, DeVincenzo R, Bonanno G, Ferrandina G, Piantelli M, Bussa S, Rumi C, Cianfriglia M, Mancuso S (1994) Quercetin potentiates the effect of adriamycin in a multidrug-resistant MCF-7 human breast cancer cell line: P-glycoprotein as a possible target. Cancer Chemother Pharmacol 34:459–464CrossRefPubMedGoogle Scholar
  29. 29.
    Boots AW, Li H, Schins RP, Duffin R, Heemskerk JW, Bast A, Haenen GR (2007) The quercetin paradox. Toxicol Appl Pharmacol 222:89–96CrossRefPubMedGoogle Scholar
  30. 30.
    Egert S, Wolffram S, Bosy-Westphal A, Boesch-Saadatmandi C, Wagner AE, Frank J, Rimbach G, Mueller MJ (2008) Daily quercetin supplementation dose-dependently increases plasma quercetin concentrations in healthy humans. J Nutr 138:1615–1621PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Gangjun Du
    • 1
    • 2
  • Haihong Lin
    • 1
  • Mei Wang
    • 1
  • Shuo Zhang
    • 1
    • 2
  • Xianchuang Wu
    • 1
  • Linlin Lu
    • 1
  • Liyan Ji
    • 1
  • Lijuan Yu
    • 1
  1. 1.Department of PharmacologyPharmaceutical College of Henan UniversityKaifengChina
  2. 2.Protein and Peptide Pharmaceutical Laboratory, Institute of BiophysicsChinese Academy of SciencesBeijingChina

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