Skip to main content
Log in

Arsenic induces DNA damage via reactive oxygen species in human cells

  • Original Article
  • Published:
Environmental Health and Preventive Medicine Aims and scope

Abstract

To elucidate arsenic-induced oxidative DNA damage, the genotoxicity of arsenic in human cells was comparatively studied with single cell gel electrophoresis (SCGE) assay in combination with the observation of the protective effects of dimethyl sulfoxide (DMSO) and catalase. Arsenic, at the concentration of 2.4 μM by coincubation for 24 hours, significantly induced DNA damage in HL60, a human promyelocytic leukemia cell line. In contrast, significant DNA damage was found in human mononucleocytes at the concentration of 4.8 μM or above. The cells were incubated separately with DMSO (12 mM/l), a well-known hydroxyl radical (OH) scavenger, and catalase (1,300 U/ml), a hydrogen peroxide (H2O2) scavenger, for 6 hours and then further coincubated with various concentrations of arsenic for 24 hours at 37°C and 5% CO2. The findings showed that both DMSO and catalase significantly reduced the arsenic-induced tail moment, a parameter of total damaged DNA, in HL60 and mononucleocytes. Hence our findings indicate that arsenic, with micromolar concentrations, induces typical and various extents of DNA damage in human cells via reactive oxygen species in a dose-dependent manner.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Rossman T, Thompson C and Waalkes M Arsenic: health effects, mechanisms of actions, and research issues. Environ. Health Perspect. 1999; 107: 593–597.

    Article  PubMed  Google Scholar 

  2. Lee-Chen SF, Gurr JR, Lin JB and Jan KY. Arsenite enhances DNA double-strand breaks and cell-killing of methyl methane-sulfonate-treated cells by inhibiting the excision of alkali-labile sites. Mutat. Res. 1993; 294: 21–27.

    PubMed  CAS  Google Scholar 

  3. Lynn S, Lai HT, Gurr JR and Jan KY. Arsenite retards DNA break rejoining by inhibiting DNA ligation. Mutagenesis. 1997; 12: 353–358.

    Article  PubMed  CAS  Google Scholar 

  4. Yager JW and Wiencke K. Inhibition of poly (ADP-ribose) polymerase by arsenite. Mutat. Res. 1997; 386: 345–346.

    Article  PubMed  CAS  Google Scholar 

  5. Hei TK, Liu SX and Waldren C. Mutagenicity of arsenic in mammalian cells: role of reactive oxygen species. Proc. Natl. Acad. Sci. USA 1998; 95: 8103–8104.

    Article  PubMed  CAS  Google Scholar 

  6. Wang TS, Shu YF, Liu YC, Jan KY and Huang H. Glutathione peroxidase and catalase modulate the genotoxicity of arsenite. Toxicology 1997; 121: 229–238.

    Article  PubMed  CAS  Google Scholar 

  7. Matsui M, Nishigori C, Toyokuni S, Takada J, Akaboshi M, Ishikawa M, Imamura S and Miyachi Y. The role of oxidative DNA damage in human arsenic carcinogenesis: detection of 8-hydroxy-2′-deoxyguanosine in arsenic-related Bowen’s disease. J. Invest. Dermatol. 1999; 113: 26–35.

    Article  PubMed  CAS  Google Scholar 

  8. Dong JT and Luo XM. Arsenic-induced DNA-strand breaks associated with DNA-protein crosslinks in human fetal lung fibroblasts. Mutat. Res. 1993; 302: 97–105.

    Article  PubMed  CAS  Google Scholar 

  9. Singh NP, McCoy MT, Tice RR and Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp. Cell Res. 1988; 175: 184–187.

    Article  PubMed  CAS  Google Scholar 

  10. Lu Y, Takeshita T and Morimoto K. Single-cell gel electrophoresis (SCG)-A review and discussion. Environ. Health Prev. Med. 1997; 2: 53–55.

    Article  CAS  Google Scholar 

  11. Kobayashi H and Hayashi M. Classification and evaluation of comets in single cell gel electrophoresis assay. Environ. Mutagen Res. 1999; 21: 231–236.

    CAS  Google Scholar 

  12. Heydorn K. Environmental variation of arsenic levels in human blood determines by neutron activation analysis. Clin. Chim. Acta 1970; 28: 349–356.

    Article  PubMed  CAS  Google Scholar 

  13. Kinnula VL, Whorton AR, Chang LY and Crapo JD. Regulation of hydrogen peroxide generation in cultured endothelial cells. Am. J. Respir. Cell Mol. Biol. 1992; 6: 175–177.

    PubMed  CAS  Google Scholar 

  14. Kobzik L, Godleski JJ and Brain JD. Oxidative metabolism in the alveolar macrophage: analysis by flow cytometry. J. Leukoc. Biol. 1990; 47: 295–298.

    PubMed  CAS  Google Scholar 

  15. Carter WO, Narayanan PK and Robinson JP. Intracellular hydrogen peroxide and superoxide anion detection in endothelial cells. J. Leukoc. Biol. 1994; 55: 253–258.

    PubMed  CAS  Google Scholar 

  16. Sarker AH, Watanabe S, Seki S, Akiyama T and Okada S. Oxygen radical-induced single-strand DNA breaks and repair of the damage in a cell-free system. Mutat. Res. 1995; 337: 85–110.

    PubMed  CAS  Google Scholar 

  17. Lee TC and Ho IC. Modulation of cellular antioxidant defense activities by sodium arsenite in human fibroblasts. Arch. Toxicol. 1995; 67: 498–506.

    Article  Google Scholar 

  18. Wang TS, Kuo CF, Jan KY and Huang H. Arsenite induces apoptosis in Chinese hamster ovary cells by generation of reactive oxygen species. J. Cell Physiol. 1996; 256–312.

  19. Barchowsky A, Klei LR, Dudek EJ, Swarts HM and James PE. Stimulation of reactive oxygen, but not reactive nitrogen species, in vascular endothelial cells exposed to low levels of arsenite. Free Radic. Biol. Med. 1999; 27: 1405–1412.

    CAS  Google Scholar 

  20. Milligan JR, Aguilera JA, Nguyen TT, Ward JF, Kow YW, He B and Cunningham RP. Yield of DNA strand breaks after base oxidation of plasmid DNA. Radiat. Res. 1999; 151: 334–338.

    Article  PubMed  CAS  Google Scholar 

  21. Henle ES and Linn S. Formation, prevention and repair of DNA damage by ion/hydrogen peroxide. J. Biol. Chem. 1997; 272: 19095–19103.

    Article  PubMed  CAS  Google Scholar 

  22. Hemnani T and Parihar MS. Reactive oxygen species and oxidative DNA damage. Indian J. Physiol. Pharmacol. 1998; 42: 440–452.

    PubMed  CAS  Google Scholar 

  23. Imlay JA and Keyer K. Superoxide accelerates DNA damage by elevating free-ion levels. Proc. Natl. Acad. Sci. USA 1996; 93: 13635–13645.

    Article  PubMed  Google Scholar 

  24. Jing Y, Dai J, Chalmers-Redman RM, Tatton WG and Waxman S. Arsenic trioxide selectively induces acute promyelocytic leukemia cell apoptosis via a hydrogen peroxide-dependent pathway. Blood 1999; 94: 2102–2111.

    PubMed  CAS  Google Scholar 

  25. Lynn S, Gurr JR, Lai HT and Jan KY. NADH oxidase activation is involved in arsenite-induced oxidative DNA damage in human vascular smooth muscle cells. Circ. Res. 2000; 86: 514–519.

    PubMed  CAS  Google Scholar 

  26. Chen YC, Lin-Shiau SY and Lin JK. Involvement of reactive oxygen species and caspase 3 activation in arsenite-induced apoptosis. J. Cell Physiol. 1998; 177: 324–329.

    Article  PubMed  CAS  Google Scholar 

  27. Collins AR, Ma AG and Duthie SJ. The kinetics of repair of oxidative DNA damage (strand breaks and oxidised pyrimidines) in human cells. Mutat. Res. 1995; 336: 69–78.

    PubMed  CAS  Google Scholar 

  28. Green MHL, Lowe JE, Delaney CA and Green IC. Comet assay to detect nitric oxide-dependent DNA damage in mammalian cells. Methods Enzymol. 1996; 269: 243–313.

    PubMed  CAS  Google Scholar 

  29. Ohyama H and Yamada T. Teardrop assay: A detection method for individual apoptotic cells with DNA fragmentation. Environ. Mutagen Res. 1999; 21: 237–244.

    Google Scholar 

  30. Wang ZG, Rivi R, Delva L, Konig A, Scheinberg DA, Gambacorti-asserini C, Gabrilove JL, Warrell RP Jr and Pandolfi PP. Arsenic trioxide and melarsoprol induce programmed cell death in myeloid leukemia cell lines and function in a PML and PML-RARalpha independent manner. Blood 1998; 92: 1497–1507.

    PubMed  CAS  Google Scholar 

  31. Marsteinstredet R, Wiger R, Brunborg G, Hongslo JK and Holme JA. Apoptosis in HL-60 cells induced by 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX). Chem. Biol. Interact. 1997; 106: 89–118.

    Article  PubMed  CAS  Google Scholar 

  32. Morris EJ, Dreixler JC, Cheng KY, Wilson PM, Gin RM and Geller HM. Optimization of single-cell gel electrophoresis (SCGE) for quantitative analysis of neuronal DNA damage. Biotechniques 1999; 26: 286–289.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Kanehisa Morimoto.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, D., Morimoto, K., Takeshita, T. et al. Arsenic induces DNA damage via reactive oxygen species in human cells. Environ Health Prev Med 6, 27–32 (2001). https://doi.org/10.1007/BF02897306

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02897306

Key words

Navigation