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Effects of ascorbic acid and β-carotene on HepG2 human hepatocellular carcinoma cell line

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Abstract

Recent studies have demonstrated that vegetable rich diets have protective effects on the occurrence and prognosis of various cancers. In addition to dietary intakes, ascorbic acid and β-carotene are also taken as supplements. The aim of this study was to assess effects of ascorbic acid, β-carotene and their combinations on human hepatocellular carcinoma cell line HepG2. Ascorbic acid and β-carotene were applied to cells as plasma peak concentrations (70 and 8 μM, respectively) and their half concentrations (35 and 4 μM, respectively) for 24 and 48 h. Genotoxic and cytotoxic effects of ascorbic acid and β-carotene were evaluated by alkali single cell gel electrophoresis (SCGE), acridine orange/ethidium bromide staining patterns of cells (apoptosis and necrosis) and lipid peroxidation (thiobarbituric acid reactive substances, TBARS). Results of the SCGE demonstrated that both ascorbic acid and β-carotene caused DNA damage on HepG2 which were also concordant to increased apoptosis and necrosis of cells. Increased TBARS values also demonstrated increased lipid peroxidation in these cells. Results of the present study demonstrates that when dietary intakes of ascorbic acid and β-carotene and their relevant achievable plasma level concentrations were considered, both ascorbic acid and β-carotene induce genotoxic and cytotoxic damage on HepG2 together with increased oxidative damage in contrast to their protective effect on healthy cells. This may be correlated to oxidative status and balance of ROS in hepatocellular carcinoma cells.

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References

  1. Woods JA, Bilton RF, Young AJ (1999) Beta carotene enhances hydrogen peroxide-induced DNA damage in human hepatocellular HepG2 cells. FEBS Lett 23:255–258

    Article  Google Scholar 

  2. Bjelakovic G, Nikolova D, Simonetti RG, Gluud C (2004) Antioxidant supplements for prevention of gastrointestinal cancers: a systematic review and meta-analysis. Lancet 2:1219–1228

    Article  Google Scholar 

  3. Zhang P, Omaye ST (2001) DNA strand breakage and oxygen tension: effects of β-carotene, alpha-tocopherol and AsA. Food Chem Toxicol 39:239–246

    Article  PubMed  CAS  Google Scholar 

  4. Neuhouser ML (2004) Dietary flavonoids and cancer risk: evidence from human population studies. Nutr Cancer 50:1–7

    Article  PubMed  CAS  Google Scholar 

  5. Lin J, Cook NR, Albert C, Zaharris E, Gaziano JM, Van Denburgh M et al (2009) Vitamins C and E and β carotene supplementation and cancer risk: a randomized controlled trial. J Natl Cancer Inst 7:14–23

    Google Scholar 

  6. Arranz N, Haza AI, García A, Rafter J, Morales P (2007) Protective effect of vitamin C towards N-nitrosamine-induced DNA damage in the single-cell gel electrophoresis (SCGE)/HepG2 assay. Toxicol In Vitro 21:1311–1317

    Article  PubMed  CAS  Google Scholar 

  7. Dias CD, Araújo BC, Dutra ES, Nepomuceno JC (2009) Protective effects of beta carotene against the genotoxicity of doxorubicin in somatic cells of Drosophila melanogaster. Genet Mol Res 17:1367–1375

    Article  Google Scholar 

  8. O’Brien NM, Woods JA, Aherne SA, O’Callaghan YC (2000) Cytotoxicity, genotoxicity and oxidative reactions in cell-culture models: modulatory effects of phytochemicals. Biochem Soc Trans 28:22–26

    PubMed  Google Scholar 

  9. VanderJagt DJ, Garry PJ, Bhagavan HN (1987) Ascorbic acid intake and plasma levels in healthy elderly people. Am J Clin Nutr 46:290–294

    PubMed  CAS  Google Scholar 

  10. Micozzi MS, Brown ED, Edwards BK, Bieri JG, Taylor PR, Khachik F et al (1992) Plasma carotenoid response to chronic intake of selected foods and BC supplements in men. Am J Clin Nutr 55:1120–1125

    PubMed  CAS  Google Scholar 

  11. Baskić D, Popović S, Ristić P, Arsenijević NN (2006) Analysis of cycloheximide-induced apoptosis in human leukocytes: fluorescence microscopy using annexin V/propidium iodide versus acridin orange/ethidium bromide. Cell Biol Int 30:924–932

    Article  PubMed  Google Scholar 

  12. Ahn DU, Olson DG, Jo C, Chen X, Cand Wu, Lee JI (1998) Effect of muscle type, packaging, and irradiation on lipid oxidation, volatile production, and color in raw pork patties. Meat Sci 49:27–39

    Article  CAS  Google Scholar 

  13. Bradford MM (1976) Rapid and sensitive method for quantitation of microgram quantities of protein utilizing principle of protein–dye binding. Anal Biochem 72:248–254

    Article  PubMed  CAS  Google Scholar 

  14. García A, Morales P, Arranz N, Delgado ME, Rafter J, Haza AI (2009) Antiapoptotic effects of dietary antioxidants towards N-nitrosopiperidine and N-nitrosodibutylamine-induced apoptosis in HL-60 and HepG2 cells. J Appl Toxicol 29:403–413

    Article  PubMed  Google Scholar 

  15. Prakash P, Russell RM, Krinsky NI (2001) In vitro inhibition of proliferation of estrogen-dependent and estrogen-independent human breast cancer cells treated with carotenoids or retinoids. J Nutr 131:1574–1580

    PubMed  CAS  Google Scholar 

  16. Li JJ, Tang Q, Li Y, Hu BR, Ming ZY, Fu Q et al (2006) Role of oxidative stress in the apoptosis of hepatocellular carcinoma induced by combination of arsenic trioxide and ascorbic acid. Acta Pharmacol Sin 27:1078–1084

    Article  PubMed  CAS  Google Scholar 

  17. van Helden YG, Keijer J, Heil SG, Picó C, Palou A, Oliver P et al (2009) BC affects oxidative stress-related DNA damage in lung epithelial cells and in ferret lung. Carcinogenesis 30:2070–2076

    Article  PubMed  Google Scholar 

  18. Belin S, Kaya F, Duisit G, Giacometti S, Ciccolini J, Fontés M (2009) Antiproliferative effect of Ascorbic acid is associated with the inhibition of genes necessary to cell cycle progression. PLoS One 4:e4409

    Article  PubMed  Google Scholar 

  19. Prasad KN, Kumar A, Kochupillai V, Cole WC (1999) High doses of multiple antioxidant vitamins: essential ingredients in improving the efficacy of standard cancer therapy. J Am Coll Nutr 18:13–25

    PubMed  CAS  Google Scholar 

  20. Lee SK, Kang JS, Da Jung J, Hur DY, Kim JE, Hahm E et al (2008) Vitamin C suppresses proliferation of the human melanoma cell SK-MEL-2 through the inhibition of cyclooxygenase-2 (COX-2) expression and the modulation of insulin-like growth factor II (IGF-II) production. J Cell Physiol 216:180–188

    Article  PubMed  CAS  Google Scholar 

  21. Lowe GM, Booth LA, Young AJ, Bilton RF (1999) Lycopene and Beta Carotene protect against oxidative damage in HT29 cells at low concentrations but rapidly lose this capacity at higher doses. Free Radic Res 30:141–151

    Article  PubMed  CAS  Google Scholar 

  22. Verrax J, Calderon PB (2008) The controversial place of vitamin C in cancer treatment. Biochem Pharmacol 15:1644–1652

    Article  Google Scholar 

  23. Burton GW, Ingold KU (1984) Beta-carotene: an unusual type of lipid antioxidant. Science 11:569–573

    Article  Google Scholar 

  24. Du J, Martin SM, Levine M, Wagner BA, Buettner GR, Wang SH, Taghiyev AF, Du C, Knudson CM, Cullen JJ (2010) Mechanisms of ascorbate-induced cytotoxicity in pancreatic cancer. Clin Cancer Res 16(2):509–520

    Article  PubMed  CAS  Google Scholar 

  25. Riordan NH, Riordan HD, Meng X, Li Y, Jackson JA (1995) Intravenous ascorbate as a tumor cytotoxic chemotherapeutic agent. Med Hypotheses 44:207–213

    Article  PubMed  CAS  Google Scholar 

  26. Riordan HR, Jackson JA, Schultz M (1990) Case study: high-dose intravenous vitamin C in the treatment of a patient with adenocarcinoma of the kidney. J Ortho Med 5:5–7

    Google Scholar 

  27. Riordan HD, Jackson JA, Riordan NH, Schultz M (1998) High-dose intravenous vitamin C in the treatment of a patient with renal cell carcinoma of the kidney. J Ortho Med 13:72–73

    Google Scholar 

  28. Jackson JA, Riordan HD, Hunninghauke RE, Riordan N (1995) High dose intravenous vitamin C and long time survival of a patient with cancer of the head of the pancreas. J Ortho Med 10:87–88

    Google Scholar 

  29. Langemann H, Torhorst J, Kabiersch A, Krenger W, Honegger CG (1989) Quantitative determination of water- and lipid-soluble antioxidants in neoplastic and non-neoplastic human breast tissue. Int J Cancer 15:1169–1173

    Article  Google Scholar 

  30. Chen Q, Espey MG, Krishna MC, Mitchell JB, Corpe CP, Buettner GR, Shacter E, Levine M (2005) Pharmacologic ascorbic acid concentrations selectively kill cancer cells: action as a pro-drug to deliver hydrogen peroxide to tissues. Proc Natl Acad Sci USA 102(38):13604–13609

    Article  PubMed  CAS  Google Scholar 

  31. Casciari JJ, Riordan NH, Schmidt TL, Meng XL, Jackson JA, Riordan HD (2001) Cytotoxicity of ascorbate, lipoic acid, and other antioxidants in hollow fibre in vitro tumours. Br J Cancer 1:1544–1550

    Article  Google Scholar 

  32. Nishinoa H, Murakoshic M, Tokudad H, Satomid Y (2009) Cancer prevention by carotenoids. Arch Biochem Biophys 483(2):165–168

    Article  Google Scholar 

  33. Seifter E, Rettura A, Padawar J, Levenson SM (1984) Vitamin A and β-carotene as adjunctive therapy to tumor excision, radiation therapy and chemotherapy. In: Prasad KN (ed) Vitamins, nutrition and cancer. Karger, Basel, pp 1–19

    Google Scholar 

  34. Schwartz JL (1995) Molecular and biochemical control of tumor growth following treatment with carotenoids or tocopherols. In: Prasad KN, Santamaria L, Williams RM (eds) Nutrients in cancer prevention and treatment. Humana Press, Totawa, NJ, pp 287–316

    Chapter  Google Scholar 

  35. Shklar G (1995) Inhibition and regression of experimental oral cancer by beta carotene and vitamin E: emerging concepts. In: Prasad KN, Santamaria L, Williams RM (eds) Nutrients in cancer prevention and treatment. Humana Press, Totowa, NJ, pp 317–333

    Chapter  Google Scholar 

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Acknowledgments

This study was approved by Baskent University Institutional Review Board (Project No. DA10/01) and supported by Baskent University Research Fund.

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Correspondence to Feride I. Sahin.

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Yurtcu, E., Iseri, O.D. & Sahin, F.I. Effects of ascorbic acid and β-carotene on HepG2 human hepatocellular carcinoma cell line. Mol Biol Rep 38, 4265–4272 (2011). https://doi.org/10.1007/s11033-010-0549-5

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  • DOI: https://doi.org/10.1007/s11033-010-0549-5

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