Tumor Biology

, Volume 36, Issue 12, pp 9267–9275 | Cite as

Effect of hesperidin on mice bearing Ehrlich solid carcinoma maintained on doxorubicin

  • Naglaa F. Khedr
  • Rania M. Khalil
Research Article


Doxorubicin (DOX) is widely used in cancer therapy of many carcinomas types. Unfortunately, DOX is not sufficiently effective in many cases, and increasing the dosage of it is limited due to its systemic toxicity. A citrus flavonoid hesperidin (HES) is proved to be potent antioxidant and protective agent against many diseases including cancer. In this context, the objective of this study was to examine effect of HES along with DOX on solid Ehrlich carcinoma (SEC) in mice. Forty male mice were divided into four equal groups (n = 10): control SEC, DOX, HES, and DOX + HES. HES (50 mg/kg body weight orally) was given day after day for 16 days along with DOX (4 mg/kg body weight i.p. injection) for 5 cycles every 4 days in ESC-inoculated mice. After 20 days, tumor volume, tumor weight, survival rate, tumor glutathione, nitric oxide content, and serum glutathione were determined. Tumor tissue was examined for histopathological and immunohistochemical study for p53 and VEGF. Tumor resistance for mdr1a gene was assessed in tumor tissue by RT-PCR. HES induced significant increase in tissue and serum glutathione with significant decrease in tumor volume and tumor weight. A possible role of HES to modulate gene expression of mdr1a in tumor tissue was established. In addition, HES alleviated the histopathological changes with significant decrease in p53 and VEGF expression. The use of HES as adjuvant therapy with DOX would enhance the therapeutic efficacy and alleviate the resistance to DOX in treatment of solid tumors.


Doxorubicin Ehrlich solid tumor Flavonoids Glutathione Hesperidin P53 VEGF 


  1. 1.
    Agrawal SS, Saraswati S, Mathur R, Pandey M. Cytotoxic and antitumor effects of brucine on Ehrlich ascites tumor and human cancer cell line. Life Sci. 2011;89:147–58.CrossRefPubMedGoogle Scholar
  2. 2.
    Alamed J, Chaiyasit W, McClements DJ, Decker EA. Relationships between free radical scavenging and antioxidant activity in foods. J Agric Food Chem. 2009;57(7):2969–76.CrossRefPubMedGoogle Scholar
  3. 3.
    Arafa HM, Aly HA, Abd-Ellah MF, El-Refaey HM. Hesperidin attenuates benzo[alpha] pyrene-induced testicular toxicity in rats via regulation of oxidant/antioxidant balance. Toxicol Ind Health. 2009;25(6):417–27.CrossRefPubMedGoogle Scholar
  4. 4.
    Bustin and Stephen A. (ed) A-Z of Quantitative PCR. La Jolla, CA: International University Line. 2004–2006.Google Scholar
  5. 5.
    Chatterjee K, Zhang J, Honbo N, Karliner JS. Doxorubicin cardiomyopathy. Cardiology. 2010;115(2):155–62.CrossRefPubMedGoogle Scholar
  6. 6.
    Choi EJ, Kim GH. Anti-/pro-apoptotic effects of hesperetin against 7,12-dimetylbenz(a)anthracene-induced alteration in animals. Oncol Rep. 2011;25(2):545–50.CrossRefPubMedGoogle Scholar
  7. 7.
    Corpet DE, Taché S. Ranking chemopreventive agents on rat colon carcinogenesis. IARC Sci Publ. 2002;156:381–4.PubMedGoogle Scholar
  8. 8.
    Dabkeviciene D, Sasnauskiene A, Leman E, Kvietkauskaite R, Kirveliene V. Differential expression of VEGF and IL-1alpha after photodynamic treatment in combination with doxorubicin or taxotere. Anticancer Res. 2014;34(10):5295–302.PubMedGoogle Scholar
  9. 9.
    Darvesh AS, Aggarwal BB, Bishayee A. Curcumin and liver cancer: a review. Curr Pharm Biotechnol. 2012;13(1):218–28.CrossRefPubMedGoogle Scholar
  10. 10.
    Deavall DG, Martin EA, Horner JM, Roberts R. Drug-induced oxidative stress and toxicity. J Toxicol. 2012;645460–72.Google Scholar
  11. 11.
    El-Ashmawy N, Khalil R. A review on the role of L-carnitine in the management of tamoxifen side effects in treated women with breast cancer. Tumor Biol. 2014;35(4):2845–55.CrossRefGoogle Scholar
  12. 12.
    Ellman GL. Tissue Sulfhydryl Groups. Arch Biochem Biophys. 1959;82:70–7.CrossRefPubMedGoogle Scholar
  13. 13.
    Erbas H, Aydogdu N, Usta U, Erten O. Protective role of carnitine in breast cancer via decreasing arginase activity and increasing nitric oxide. Cell Biol Int. 2007;31:1414–9.CrossRefPubMedGoogle Scholar
  14. 14.
    Firuzi O, Miri R, Tavakkoli M, Saso L. Antioxidant therapy: current status and future prospects. Curr Med Chem. 2011;18(25):3871–88.CrossRefPubMedGoogle Scholar
  15. 15.
    Gammella E, Maccarinelli F, Buratti P, Recalcati S, Cairo G. The role of iron in anthracycline cardiotoxicity. Front Pharmacol. 2014;26:5–25.Google Scholar
  16. 16.
    Gao JZ, Wang YL, Li J. i LX. Effects of VEGF/VEGFR/K-ras signaling pathways on miRNA21 levels in hepatocellular carcinoma tissues in rats. Genet Mol Res. 2015;14(1):671–9.CrossRefPubMedGoogle Scholar
  17. 17.
    Guan J, Chen J. Mesenchymal stem cells in the tumor microenvironment. Biomed Rep. 2013;1(4):517–21.PubMedPubMedCentralGoogle Scholar
  18. 18.
    Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell. 2011;144(5):646–74.CrossRefPubMedGoogle Scholar
  19. 19.
    Hwang SL, Yen GC. Effect of hesperetin against oxidative stress via ER- and TrkA-mediated actions in PC12 cells. J Agric Food Chem. 2011;59(10):5779–85.CrossRefPubMedGoogle Scholar
  20. 20.
    Jäeger W. Classical resistance mechanisms. Int J Clin Pharmacol Ther. 2009;47(1):46–8.CrossRefPubMedGoogle Scholar
  21. 21.
    Jaganathan SK, Mondhe D, Wani ZA, Pal HC, Mandal M. Effect of honey and eugenol on Ehrlich ascites and solid carcinoma. J Biomed Biotechnol. 2010;989163–67.Google Scholar
  22. 22.
    Kamaraj S, Ramakrishnan G, Anandakumar P, Jagan S, Devaki T. Antioxidant and anticancer efficacy of hesperidin in benzo(a)pyrene induced lung carcinogenesis in mice. Invest New Drugs. 2009;27(3):214–22.CrossRefPubMedGoogle Scholar
  23. 23.
    L’Ecuyer T, Sanjeev S, Thomas R, Novak R, Das L, Campbell W, et al. DNA damage is an early event in doxorubicin-induced cardiac myocyte death. Am J Physiol Heart Circ Physiol. 2006;291(3):H1273–80.CrossRefPubMedGoogle Scholar
  24. 24.
    Li D, Li J, An Y, Yang Y, Zhang SQ. Doxorubicin-induced apoptosis in H9c2 cardiomyocytes by NF-κB dependent PUMA upregulation. Eur Rev Med Pharmacol Sci. 2013;17(17):2323–9.PubMedGoogle Scholar
  25. 25.
    Liu J, Zhang C, Feng Z. Tumor suppressor p53 and its gain-of-function mutants in cancer. Acta Biochim Biophys Sin (Shanghai). 2014;46(3):170–9.CrossRefGoogle Scholar
  26. 26.
    Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin- phenol reagent. J Biol Chem. 1951;193:265–75.PubMedGoogle Scholar
  27. 27.
    Maciag AE, Chakrapani H, Saavedra JE, Morris NL, Holland RJ, Kosak KM, et al. The nitric oxide prodrug JS-K is effective against non-small-cell lung cancer cells in vitro and in vivo: involvement of reactive oxygen species. J Pharmacol Exp Ther. 2011;336(2):313–20.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Manne U, Weiss HL, Myers RB, Danner OK. Nuclear accumulation of p53 in colorectal adenocarcinomas: prognostic importance differs with race and location of tumor. Cancer. 1998;83:2456–67.CrossRefPubMedGoogle Scholar
  29. 29.
    Martinez L, Arnaud O, Henin E, Tao H, Chaptal V, Doshi R, et al. Understanding polyspecificity within the substrate-binding cavity of the human multidrug resistance P-glycoprotein. FEBS J. 2014;281(3):673–82.CrossRefPubMedGoogle Scholar
  30. 30.
    Meiyanto E, Hermawan A, Anindyajati. Natural products for cancer-targeted therapy: citrus flavonoids as potent chemopreventive agents. Asian Pac J Cancer Prev. 2012;13(2):427–36.CrossRefPubMedGoogle Scholar
  31. 31.
    Menendez D, Nguyen TA, Freudenberg JM, Mathew VJ, Anderson CW, Jothi R, et al. Diverse stresses dramatically alter genome-wide p53 binding and transactivation landscape in human cancer cells. Nucleic Acids Res. 2013;41(15):7286–301.CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Miranda-Vilela AL, Portilho FA, de Araujo VG, Estevanato LL, Mezzomo BP, Santos Mde F, et al. The protective effects of nutritional antioxidant therapy on Ehrlich solid tumor bearing mice depend on the type of antioxidant therapy chosen: histology, genotoxicity and hematology evaluations. J Nutr Biochem. 2011;22(11):1091–8.CrossRefPubMedGoogle Scholar
  33. 33.
    Mizutani T, Hattori A. New horizon of MDR1 (P-glycoprotein) study. Drug Metab Rev. 2005;37(3):489–510.CrossRefPubMedGoogle Scholar
  34. 34.
    O’Connor R, Clynes M, Dowling P, O’Donovan N, O’Driscoll L. Drug resistance in cancer - searching for mechanisms, markers and therapeutic agents. Expert Opin Drug Metab Toxicol. 2007;3(6):805–17.CrossRefPubMedGoogle Scholar
  35. 35.
    Oz E, Ilhan MN. Effects of melatonin in reducing the toxic effects of doxorubicin. Mol Cell Biochem. 2006;286(1–2):11–5.CrossRefPubMedGoogle Scholar
  36. 36.
    Pari L, Karthikeyan A, Karthika P, Rathinam A. Protective effects of hesperidin on oxidative stress, dyslipidaemia and histological changes in iron-induced hepatic and renal toxicity in rats. Toxicol Rep. 2015;2:46–55.CrossRefGoogle Scholar
  37. 37.
    Park HJ, Kim MJ, Ha E, Chung JH. Apoptotic effect of hesperidin through caspase3 activation in human colon cancer cells, SNU-C4. Phytomedecine. 2008;15(1–2):147–51.CrossRefGoogle Scholar
  38. 38.
    Ridnour LA, Sim JE, Hayward MA, Wink DA, Martin SM, Buettner GR, et al. A spectrophotometric method for the direct detection and quantitation of nitric oxide, nitrite, and nitrate in cell culture media. Anal Biochem. 2000;281(2):223–9.CrossRefPubMedGoogle Scholar
  39. 39.
    Ross JA, Kasum CM. Dietary flavonoids: bioavailability, metabolic effects, and safety. Annu Rev Nutr. 2002;22:19–34.CrossRefPubMedGoogle Scholar
  40. 40.
    Sakata K, Hirose Y, Qiao Z, Tanaka T, Mori H. Inhibition of inducible isoforms of cyclooxygenase and nitric oxide synthase by flavonoid hesperidin in mouse macrophage cell line. Cancer Lett. 2003;199(2):139–45.CrossRefPubMedGoogle Scholar
  41. 41.
    Zhang FY, Du GJ, Zhang L, Zhang CL, Lu WL, Liang W. Naringenin enhances the anti-tumor effect of doxorubicin through selectively inhibiting the activity of multidrug resistance-associated proteins but not P-glycoprotein. Pharm Res. 2009;26:914–25.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  1. 1.Biochemistry Department, Faculty of PharmacyTanta UniversityTantaEgypt
  2. 2.Biochemistry Department, Faculty of PharmacyAL-Delta UniversityGamasaEgypt

Personalised recommendations