Hostname: page-component-76fb5796d-2lccl Total loading time: 0 Render date: 2024-04-25T11:52:39.638Z Has data issue: false hasContentIssue false
  • English
  • Français

DNA damage and susceptibility to oxidative damage in lymphocytes: effects of carotenoids in vitro and in vivo

Published online by Cambridge University Press:  09 March 2007

Siân B. Astley ,
David A. Hughes ,
Anthony J. A. Wright ,
Ruan M. Elliott  and
Susan Southon
Siân B. Astley*
Affiliation:
Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, UK
David A. Hughes
Affiliation:
Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, UK
Anthony J. A. Wright
Affiliation:
Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, UK
Ruan M. Elliott
Affiliation:
Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, UK
Susan Southon
Affiliation:
Institute of Food Research, Norwich Research Park, Colney, Norwich NR4 7UA, UK
*
*Corresponding author: Dr Siân B. Astley, fax +44 1603 255 167, email sian.astley@bbsrc.ac.uk
Rights & Permissions [Opens in a new window]

Abstract

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

Reports on the effects of carotenoids are conflicting. The present paper examines similarities and differences from contiguous studies in vitro and in vivo. Single-cell gel electrophoresis was used to measure the frequency of single-strand breaks (SSB) in the cell line MOLT-17 (as a model system) and human peripheral blood lymphocytes (PBL). MOLT-17 cells were supplemented with β-carotene, lutein or lycopene at a range of concentrations (0·00–8·00 μmol/l) using a liposome delivery method. Uptake was dose-dependent. β-Carotene concentration in the media had no effect on SSB in control cells, but incubation with lycopene or lutein (>2·00 μmol/l) increased the numbers of SSB in control cells. MOLT-17 DNA was less susceptible to oxidative damage (100 μmol H2O2/l, 5 min, 4 °C) following incubation with carotenoids between 0·50 and 1·00 μmol/l; at >1·00 μmol/l the effects were ambiguous. Apparently healthy male volunteers supplemented their habitual diets with lutein, β-carotene or lycopene (natural isolate capsules, 15 mg/d, 4 weeks) in three independent studies, raising plasma concentrations to different extents. Lycopene and lutein had no effect on SSB in control PBL or following oxidative challenge. However, increased plasma β-carotene was associated with more SSB in control cells whilst PBL DNA resistance to oxidative damage ex vivo was unaffected. These results suggest that the carotenoids are capable of exerting two overlapping but distinct effects: antioxidant protection by scavenging DNA-damaging free radicals and modulation of DNA repair mechanisms.

Keywords

CarotenoidsDNA damage and repairSingle-cell gel electrophoresisPeripheral blood lymphocytes
Type
Research Article
Information
British Journal of Nutrition , Volume 91 , Issue 1 , January 2004 , pp. 53 - 61
Copyright
Copyright © The Nutrition Society 2004

References

Astley, SB (1997) Limitation of DNA damage: the role of carotenoids and alpha-tocopherol from the diet, PhD thesis, University of East Anglia.Google Scholar
Astley, SB, Elliott, RM, Archer, DB & Southon, S (2002) Increased cellular carotenoid levels reduce the persistence of DNA single-strand breaks after oxidative challenge. Nutr Cancer 43, 202–213.CrossRefGoogle ScholarPubMed
Astley, SB, Elliott, RM, Archer, DB & Southon, S (2003) Evidence that dietary supplementation with carotenoids and carotenoid-rich foods modulates the DNA damage: repair balance in human lymphocytes. Br J Nut (In the Press).Google Scholar
Astley, SB, Hughes, DA, Wright, AJA, Peerless, ACJ & Southon, S (1996) Effect of beta-carotene supplementation on DNA damage in human blood lymphocytes. Biochem Soc Trans 24, 526S.CrossRefGoogle Scholar
Astley, SB, Pinder, AC & Southon, S (1994) DNA damage in cultures human T-lymphocytic cells as detected by the comet assay: effect of increased cellular beta-carotene and alpha-tocopherol. Proc Nutr Soc 53, 140A.Google Scholar
Collins, AR, Ma, AG & Duthie, SJ (1995) The kinetics of repair of oxidative DNA damage (strand breaks and oxidised pyrimidines) in human cells. Mutat Res 336, 69–77.CrossRefGoogle ScholarPubMed
Collins, AR, Olmedilla, B, Southon, S, Granado, F & Duthie, SJ (1998) Serum carotenoids and oxidative DNA damage in human lymphocytes. Carcinogenesis 19, 2159–2162.CrossRefGoogle ScholarPubMed
Cooke, MS, Evans, MD, Podmore, ID, et al. (1998) Novel repair action of vitamin C upon in vivo oxidative DNA damage. FEBS Letters 439, 363–367.CrossRefGoogle ScholarPubMed
Cooke, MS, Podmore, ID, Evans, MD & Lunec, J (2001) Antioxidant properties of vitamin C: new issues. Cancer Detection Prev 25, 565–567.Google ScholarPubMed
Cooke, MS, Evans, MD, Mistry, N & Lunec, J (2002) Role of dietary antioxidants in the prevention of in vivo oxidative DNA damage. Nutr Res Rev 15, 19–41.CrossRefGoogle ScholarPubMed
Drexler, HG & Minowada, J (1989) Morphological, immunophenotypical and isoenzymatic profiles of human leukemia cells and derived T-cell lines. Hematol Oncol 7, 115–125.CrossRefGoogle ScholarPubMed
Dušinška, M, Vallová, B, Ursínyová, M, et al. (2002) DNA damage and antioxidants; fluctuations through the year in a central European population group. Food Chem Toxicol 40, 1119–1123.CrossRefGoogle Scholar
Duthie, SJ, Ma, A, Ross, MA & Collins, AR (1996) Antioxidant supplementation decreases oxidative DNA damage in human lymphocytes. Cancer Res 56, 1291–1295.Google ScholarPubMed
Fillion, L, Collins, A & Southon, S (1998) Beta-carotene enhances the recovery of lymphocytes from oxidative DNA damage. Acta Biochim Pol 45, 183–190.CrossRefGoogle ScholarPubMed
Grolier, P, Azais-Braesco, V, Zelmire, L & Fessi, H (1992) Incorporation of carotenoids in aqueous systems: uptake by cultured rat hepatocytes. Biochim Biophys Acta 1111, 135–138.CrossRefGoogle ScholarPubMed
Hart, DJ & Scott, KJ (1995) Development and evaluation of an HPLC method for the analysis of carotenoids in foods, and the measurement of the carotenoid content of vegetables and fruit comonly consumed in the UK. Food Chem 54, 101–111.CrossRefGoogle Scholar
het Hof, KH, Gartner, C, Wiersma, A, Tijburg, LB & Weststrate, JA (1999) Comparison of the bioavailability of natural palm oil carotenoids and synthetic beta-carotene in humans. J Agric Food Chem 47, 1582–1586.CrossRefGoogle ScholarPubMed
Izumi, T, Hazra, TK, Boldogh, I, et al. (2000) Requirement for human AP endonuclease 1 for repair of 3'-blocking damage at DNA single-strand breaks induced by reactive oxygen species. Carcinogenesis 21, 1329–1334.CrossRefGoogle Scholar
Jackson, MJ, Papa, S, Bolanos, J, et al.(%2002) Antioxidants, reactive oxygen and nitrogen species, gene induction and mitochondrial function. J Mol Aspects Med 23, 209–285.CrossRefGoogle ScholarPubMed
Jayarajan, P, Reddy, V & Mohanram, M (1980) Effect of dietary fat on absorption of beta carotene from green leafy vegetables in children. Indian J Med Res 71, 53–56.Google ScholarPubMed
Kaminskas, E & Li, JC (1992) Repair of DNA damage induced by oxygen radicals in human non-proliferating and proliferating lymphocytes. Mutat Res 274, 103–110.CrossRefGoogle ScholarPubMed
Kaplan, LA, Lau, JM & Stein, EA (1990) Carotenoid composition, concentrations, and relationships in various human organs. Clin Physiol Biochem 8, 1–10.Google ScholarPubMed
Landrum, JT & Bone, RA (2001) Lutein, zeaxanthin, and the macular pigment. Arch Biochem Biophys 385, 28–40.CrossRefGoogle ScholarPubMed
Lovell, DP, Thomas, G & Dubow, R (1999) Issues related to the experimental design and subsequent statistical analysis of in vivo and in vitro comet studies. J Teratol Carcinog Mutagen 19, 109–119.3.0.CO;2-5>CrossRefGoogle Scholar
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.CrossRefGoogle ScholarPubMed
McKelvey-Martine, VJ, Green, MHL, Schmezer, P, Pool-Zobel, BL, de Meo, MP & Collins, AR (1993) The single cell gel electrophoresis (SCGE) assay (Comet Assay) a European review. Mutat Res 288, 47–64.CrossRefGoogle Scholar
Martin, KR, Failla, ML & Smith, JC Jr (1996) Beta-carotene and lutein protect HepG2 human liver cells against oxidant-induced damage. J Nutr 126, 2098–2106.CrossRefGoogle ScholarPubMed
Norum, KR & Blomhoff, R (1992) McCollum Award Lecture, 1992: vitamin A absorption, transport, cellular uptake, and storage. Am J Clin Nutr 56, 735–744.CrossRefGoogle ScholarPubMed
Olmedilla, B, Granado, F, Southon, S, et al. (2001) Serum concentrations of carotenoids and vitamins A, E, and C in control subjects from five European countries. Br J Nutr 85, 227–238.CrossRefGoogle Scholar
Ormerod, MG (editor) (1990). Flow Cytometry: A Practical Approach, pp. 265–266. Oxford: IRL Press.Google Scholar
Parker, RS (1996) Absorption, metabolism, and transport of carotenoids. FASEB Journal 10, 542–551.CrossRefGoogle ScholarPubMed
Perrino, FW & Loeb, LA (1990) Animal cell DNA polymerases in DNA repair. Mutat Res 236, 289–300.CrossRefGoogle ScholarPubMed
Pool-Zobel, BL, Bub, A, Liegibel, UM, Treptow-Van Lishaut, S & Rechkemmer, G% (1998) Mechanisms by which vegetable consumption reduces genetic damage in humans. Cancer Epidemiol Biomarkers Prev 7, 891–899.Google ScholarPubMed
Pool-Zobel, BL, Bub, A, Muller, H, Wollowski, I &Rechkemmer, G (1997) Consumption of vegetables reduces genetic damage in humans: first results of a human intervention trial with carotenoid-rich foods. Carcinogenesis 18, 1847–1850.CrossRefGoogle ScholarPubMed
Porrini, M & Riso, P (2000) Lymphocyte lycopene concentration and DNA protection from oxidative damage is increased in women after a short period of tomato consumption. J Nutr 130, 189–192.CrossRefGoogle Scholar
Riso, P, Pinder, A, Santangelo, A & Porrini, M (1999) Does tomato consumption effectively increase the resistance of lymphocyte DNA to oxidative damage? Am J Clin Nutr 69, 712–718.CrossRefGoogle ScholarPubMed
Schmitz, HH, Poor, CL, Wellman, RB & Erdman, JW Jr (1991) Concentrations of selected carotenoids and vitamin A in human liver, kidney and lung tissue. J Nutr 121, 1613–1621.CrossRefGoogle ScholarPubMed
Singh, NP, McCoy, MT, Tice, RR & Schneider, EL (1988) A simple technique of low levels of DNA damage in individual cells. Exp Cell Res 175, 184–191.CrossRefGoogle ScholarPubMed
Stahl, W, Schwarz, W, Sundquist, AR & Sies, H (1992) cis-trans isomers of lycopene and beta-carotene in human serum and tissues. Arch Biochem Biophys 294, 173–177.CrossRefGoogle Scholar
Stahl, W, van den Berg, J, Arthur, J, et al. (2002) Bioavailability and metabolism. J Mol Aspects Med 23, 39–100.CrossRefGoogle ScholarPubMed
Thurnham, DI (1988) Do higher vitamin A requirements in men explain the differences between the sexes in plasma provitamin A carotenoids and retinol? Proc Natl Acad Sci USA 47, 181A.Google Scholar
Tice, RR, Agurell, E, Anderson, D, et al. (2000) Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen 35, 206–221.3.0.CO;2-J>CrossRefGoogle ScholarPubMed
Torbergsen, AC & Collins, AR (2000) Recovery of human lymphocytes from oxidative DNA damage; the apparent enhancement of DNA repair by carotenoids is probably simply an antioxidant effect. Eur J Nutr 39, 80–85.CrossRefGoogle ScholarPubMed
Van Loon, AA, Sonneveld, E, Hoogerbrugge, J, et al. (1993) Induction and repair of DNA single-strand breaks and DNA base damage at different cellular stages of spermatogenesis of the hamster upon in vitro exposure to ionizing radiation. Mutat Res 294, 139–148.CrossRefGoogle ScholarPubMed
Woods, JA, Bilton, RF & Young, AJ (1999) Beta-carotene enhances hydrogen peroxide-induced DNA damage in human hepatocellular HepG2 cells. FEBS Letters 449, 255–258.CrossRefGoogle ScholarPubMed
World Cancer Research Fund (1997) Food Nutrition and the Prevention of Cancer: A Global Perspective. Washington, DC: American Institute for Cancer Research.Google Scholar
Yeh, S-L & Hu, M-L (2000) Antioxidant and pro-oxidant effects of lycopene in comparison with beta-carotene on oxidant-induced damage in Hs68 cells. J Nutr Biochem 11, 548–554.CrossRefGoogle ScholarPubMed
Young, AJ & Lowe, GM (2001) Antioxidant and prooxidant properties of carotenoids. Arch Biochem Biophys 385, 20–27.CrossRefGoogle ScholarPubMed