Arsenic induces DNA damage via reactive oxygen species in human cells

  • Dasheng Li
  • Kanehisa Morimoto
  • Tatsuya Takeshita
  • Yuquan Lu
Original Article

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.

Key words

single cell gel electrophoresis (SCGE) assay DNA damage arsenic reactive oxygen species human cells 

References

  1. 1).
    Rossman T, Thompson C and Waalkes M Arsenic: health effects, mechanisms of actions, and research issues. Environ. Health Perspect. 1999; 107: 593–597.PubMedCrossRefGoogle Scholar
  2. 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.PubMedGoogle Scholar
  3. 3).
    Lynn S, Lai HT, Gurr JR and Jan KY. Arsenite retards DNA break rejoining by inhibiting DNA ligation. Mutagenesis. 1997; 12: 353–358.PubMedCrossRefGoogle Scholar
  4. 4).
    Yager JW and Wiencke K. Inhibition of poly (ADP-ribose) polymerase by arsenite. Mutat. Res. 1997; 386: 345–346.PubMedCrossRefGoogle Scholar
  5. 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.PubMedCrossRefGoogle Scholar
  6. 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.PubMedCrossRefGoogle Scholar
  7. 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.PubMedCrossRefGoogle Scholar
  8. 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.PubMedCrossRefGoogle Scholar
  9. 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.PubMedCrossRefGoogle Scholar
  10. 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.CrossRefGoogle Scholar
  11. 11).
    Kobayashi H and Hayashi M. Classification and evaluation of comets in single cell gel electrophoresis assay. Environ. Mutagen Res. 1999; 21: 231–236.Google Scholar
  12. 12).
    Heydorn K. Environmental variation of arsenic levels in human blood determines by neutron activation analysis. Clin. Chim. Acta 1970; 28: 349–356.PubMedCrossRefGoogle Scholar
  13. 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.PubMedGoogle Scholar
  14. 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.PubMedGoogle Scholar
  15. 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.PubMedGoogle Scholar
  16. 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.PubMedGoogle Scholar
  17. 17).
    Lee TC and Ho IC. Modulation of cellular antioxidant defense activities by sodium arsenite in human fibroblasts. Arch. Toxicol. 1995; 67: 498–506.CrossRefGoogle Scholar
  18. 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.Google Scholar
  19. 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.Google Scholar
  20. 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.PubMedCrossRefGoogle Scholar
  21. 21).
    Henle ES and Linn S. Formation, prevention and repair of DNA damage by ion/hydrogen peroxide. J. Biol. Chem. 1997; 272: 19095–19103.PubMedCrossRefGoogle Scholar
  22. 22).
    Hemnani T and Parihar MS. Reactive oxygen species and oxidative DNA damage. Indian J. Physiol. Pharmacol. 1998; 42: 440–452.PubMedGoogle Scholar
  23. 23).
    Imlay JA and Keyer K. Superoxide accelerates DNA damage by elevating free-ion levels. Proc. Natl. Acad. Sci. USA 1996; 93: 13635–13645.PubMedCrossRefGoogle Scholar
  24. 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.PubMedGoogle Scholar
  25. 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.PubMedGoogle Scholar
  26. 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.PubMedCrossRefGoogle Scholar
  27. 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.PubMedGoogle Scholar
  28. 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.PubMedGoogle Scholar
  29. 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. 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.PubMedGoogle Scholar
  31. 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.PubMedCrossRefGoogle Scholar
  32. 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

Copyright information

© Japanese Society of Hygiene 2001

Authors and Affiliations

  • Dasheng Li
    • 1
  • Kanehisa Morimoto
    • 1
  • Tatsuya Takeshita
    • 1
  • Yuquan Lu
    • 1
  1. 1.Department of Preventive MedicineOsaka University Graduate School of MedicineSuita, Osaka
  2. 2.Department of Social and Environmental MedicineOsaka University Graduate School of Medicine F1Suita, OsakaJapan
  3. 3.Guizhou Provincial Health and Antiepidemic StationGuiyangP.R. China

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