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Dietary polyphenols protect against N-nitrosamines and benzo(a)pyrene-induced DNA damage (strand breaks and oxidized purines/pyrimidines) in HepG2 human hepatoma cells

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Abstract

Background

Dietary polyphenols have been reported to have a variety of biological actions, including anticarcinogenic and antioxidant activities.

Aim of the study

In the present study we investigated the protective effect of dietary polyphenols against N-nitrosodimethylamine (NDMA), N-nitrosopyrrolidine (NPYR) and benzo(a)pyrene (BaP)-induced DNA damage (strand breaks and oxidized purines/pyrimidines) in HepG2 cells.

Methods

Human hepatocellular carcinoma (HepG2) cells, which retain many specialized liver functions and drug metabolizing enzyme activities, were used as in vitro model for human hepatocytes. NDMA, NPYR and BaP were employed to induce DNA damage. DNA damage (strand breaks, oxidized pyrimidines and oxidized purines) was evaluated by the alkaline single cell gel electrophoresis or comet assay.

Results

None of the polyphenols concentrations tested in presence or absence of Fpg (formamidopyrimidine-DNA glycosylase), or Endo III (Endonuclease III) caused DNA damage per se. Increasing concentrations of BaP (25–100 µM) induced a significant increase of DNA strand breaks, Fpg and Endo III sensitive sites in a dose dependent manner. Myricetin and quercetin decreased DNA strand breaks and oxidized pyrimidines induced by NDMA, but not oxidized purines. However, both flavonoids reduced oxidized pyrimidines and purines induced by NPYR. DNA strand breaks induced by NPYR were prevented by quercetin, but not by myricetin. BaP-induced DNA strand breaks and oxidized pyrimidines were strongly reduced by myricetin and quercetin, respectively. While oxidized purines induced by BaP were reduced by quercetin, myricetin had no protective effect. (+)-Catechin and (−)-epicatechin reduced DNA strand breaks, oxidized pyrimidines and oxidized purines induced by NDMA. DNA strand breaks, and oxidized purines induced by NPYR were also prevented by (+)-catechin and (−)-epicatechin, while the maximum reduction of oxidized pyrimidines was found by (+)-catechin and (−)-epicatechin at 10 µM. (+)-Catechin and (−)-epicatechin decreased also DNA strand breaks and oxidized pyrimidines but not oxidized purines induced by BaP.

Conclusions

Our results clearly indicate that polyphenols protect human derived cells against DNA strand breaks and oxidative DNA damage effects of NDMA, NPYR or BaP, three carcinogenic compounds which occur in the environment.

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Acknowledgments

This work has been supported by Grant ALI2005-01517 from the Ministerio de Educación y Ciencia (Spain) and by Grant 910177 from the Comunidad de Madrid and the Universidad Complutense (UCM). M.E. Delgado and A. García are recipients of Fellowships from the Ministerio de Educación y Ciencia and Universidad Complutense (UCM), Spain.

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Correspondence to Paloma Morales.

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Delgado, M.E., Haza, A.I., Arranz, N. et al. Dietary polyphenols protect against N-nitrosamines and benzo(a)pyrene-induced DNA damage (strand breaks and oxidized purines/pyrimidines) in HepG2 human hepatoma cells. Eur J Nutr 47, 479–490 (2008). https://doi.org/10.1007/s00394-008-0751-6

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  • DOI: https://doi.org/10.1007/s00394-008-0751-6

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