Skip to main content
SpringerLink
Log in

Generation of reactive oxygen species in the enzymatic reduction of PbCrO4 and related DNA damage

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Free radical reactions are believed to play an important role in the mechanism of Cr(VI)-induced carcinogenesis. Most studies concerning the role of free radical reactions have been limited to soluble Cr(VI). Various studies have shown that solubility is an important factor contributing to the carcinogenic potential of Cr(VI) compounds. Here, we report that reduction of insoluble PbCrO4 by glutathione reductase in the presence of NADPH as a cofactor generated hydroxyl radicals (•OH) and caused DNA damage. The •OH radicals were detected by electron spin resonance (ESR) using 5,5-dimethyl-N-oxide as a spin trap. Addition of catalase, a specific H2O2 scavenger, inhibited the •OH radical generation, indicating the involvement of H2O2 in the mechanism of Cr(VI)-induced •OH generation. Catalase reduced •OH radicals measured by electron spin resonance and reduced DNA strand breaks, indicating •OH radicals are involved in the damage measured. The H2O2 formation was measured by change in fluorescence of scopoletin in the presence of horseradish peroxidase. Molecular oxygen was used in the system as measured by oxygen consumption assay. Chelation of PbCrO4 impaired the generation of •OH radical. The results obtained from this study show that reduction of insoluble PbCrO4 by glutathione reductase/NADPH generates •OH radicals. The mechanism of •OH generation involves reduction of molecular oxygen to H2O2, which generates •OH radicals through a Fenton-like reaction. The •OH radicals generated by PbCrO4 caused DNA strand breakage.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Witmer CM, Harris R, Shupack SI: Oral bioavailability of chromium from a specific site. Environ Health Perspect 92: 105–110, 1991

    PubMed  Google Scholar 

  2. Bartlett RJ: Chromium cycling in soils and water: Links, gaps, and methods. Environ Health Perspect 92: 17–24, 1991

    PubMed  Google Scholar 

  3. De Flora S, Bagnasco M, Serra D, Zanacchi P: Genotoxicity of chromium compounds: A review. Mutat Res 238: 99–172, 1990

    PubMed  Google Scholar 

  4. Aiyar J, De Flora S, Wetterhahn KE: Reduction of chromium(VI) to chromium(V) by rat liver cytosolic and microsomal fractions: Is DT-diaphorase involved? Carcinogenesis 13: 1159–1166, 1992

    PubMed  Google Scholar 

  5. Freeman B, Lioy PJ: Exposure to chromium dust from homes in chromium surveillance project. Arch Environ Health 52: 213–226, 1997

    PubMed  Google Scholar 

  6. Hays RB: Review of occupational epidemiology of chromium chemicals and respiratory cancer. Sci Total Environ 71: 331–339, 1988

    PubMed  Google Scholar 

  7. Langard S: One hundred years of chromium and cancer: A review of epidemiological evidence and selected case reports. Am J Ind Med 17: 189–215, 1990

    PubMed  Google Scholar 

  8. Norseth T: The carcinogenicity of chromium. Environ Health Perspect 40: 121–130, 1981

    PubMed  Google Scholar 

  9. Tandon SK, Saxena DK, Gaur JS, Chandra SV: Comparative toxicity of trivalent and hexavalent chromium. Environ Res 15: 90–99, 1978

    Article  PubMed  Google Scholar 

  10. Petrilli FL, De Flora S: Toxicity and mutagenicity of hexavalent chromium on Salmonella typhimurium. Appl Environ Microbiol 33: 805–809, 1977

    PubMed  Google Scholar 

  11. Majone F, Levis AG: Chromosomal aberrations and sister-chromatid exchanges in Chinese hamster cells treated in vitro with hexavalent chromium compounds. Mutat Res 67: 231–238, 1979

    PubMed  Google Scholar 

  12. Connett PH, Wetterhahn KE: Metabolism of the carcinogenic chromate by cellular constituents. Struct Bonding 54: 93–124, 1983

    Google Scholar 

  13. Shi X, Chiu A, Chen CT, Halliwell B, Castranova V, Vallyathan V: Reduction of chromium(VI) and its relationship to carcinogenesis. J Toxicol Environ Health 2: 101–118, 1998

    Google Scholar 

  14. Shi X, Dalal NS: Chromium(V) and hydroxyl radical formation during the glutathione reductase-catalyzed reduction of chromium(VI). Biochem Biophys Res Commun 163: 627–634, 1989

    PubMed  Google Scholar 

  15. Leonard S, Wang S, Zang L, Castranova V, Vallyathan V, Shi X: Role of molecular oxygen in the generation of hydroxyl and superoxide anion radicals during enzymatic Cr(VI) reduction and its implication to Cr(VI)-induced carcinogenesis. J Environ Pathol Toxicol Oncol 19: 49–60, 2000

    PubMed  Google Scholar 

  16. Shi X, Mao Y, Knapton AD, Ding M, Rojanasakul Y, Gannett PM, Dalal NS, Liu K: Reaction of Cr(VI) with ascorbate and hydrogen peroxide generates hydroxyl radicals and causes DNA damage: Role of Cr(IV)-mediated Fenton-like reaction. Carcinogenesis 15: 2475–2478, 1994

    PubMed  Google Scholar 

  17. Ye J, Zhang X, Young HA, Mao Y, Shi X: Chromium(VI)-induced nuclear factor-κB activation in intact cells via free radical reactions. Carcinogenesis 16: 2401–2405, 1995

    PubMed  Google Scholar 

  18. Chen F, Ding M, Lu Y, Leonard SS, Vallyathan V, Castranova V, Shi X: Participation of MAP kinase p38 and IκB kinase in chromium(VI)-induced NF-κB and AP-1 activation. J Environ Pathol Toxicol Oncol 19: 231–238, 2000

    PubMed  Google Scholar 

  19. Wang S, Leonard SS, Ye J, Ding M, Shi X: The role of hydroxyl radical as a messenger in Cr(VI)-induced p53 activation. Am J Physiol 279: C868–C875, 2000

    Google Scholar 

  20. Ye J, Wang S, Leonard SS, Sun Y, Butterworth L, Antonini J, Ding M, Rojanasakul Y, Vallyathan V, Castranova V, Shi X: Role of reactive oxygen species and p53 in chromium(VI)-induced apoptosis. J Biol Chem 274: 34974–34980, 1999

    Article  PubMed  Google Scholar 

  21. Zhang Z, Leonard SS, Wang W, Vallyathan V, Castranova V, Shi X: Cr(VI) induces cell growth arrest through hydrogen-peroxide-mediated reactions. Mol Cell Biochem 222: 77–83, 2001

    Article  PubMed  Google Scholar 

  22. Ye X, Shi X: Gene expression profile in response to chromium-induced cell stress in A549 cells. Mol Cell Biochem 222: 189–197, 2001

    PubMed  Google Scholar 

  23. Hayes RB: Review of occupational epidemiology of chromium chemicals and respiratory cancer. Sci Total Environ 71: 331–339, 1988

    PubMed  Google Scholar 

  24. IARC. Chromium, Nickel and Welding. In: IARC Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to Humans. IARC, Lyon, France, 1990, 49, pp 49–256

    Google Scholar 

  25. Patierno SR, Banh D, Landolph JR: Transformation of C3H/10T1/2 mouse embryo cells to focus formation and anchorage independence by insoluble lead but not soluble calcium chromate: Relationship to mutagenesis and internalization of lead chromate particles. Cancer Res 48: 5280–5288, 1988

    PubMed  Google Scholar 

  26. Elias Z, Poirot O: Cytotoxic and neoplastic transforming effects of industrial hexavalent chromium pigments in Syrian hamster embryo cells. Carcinogenesis 10: 2043–2052, 1989

    PubMed  Google Scholar 

  27. Wise JP, Leonard JC, Patierno SR: Clastogenicity of lead chromate particles in hamster and human cells. Mutat Res 278: 69–79, 1992

    PubMed  Google Scholar 

  28. Wise JP, Stearns DM, Wetterhahn KE, Patierno SR: Cell-enhanced dissolution of carcinogenic lead chromate particles: The role of individual dissolution products in clastogenesis. Carcinogenesis 15: 2249–2254, 1994

    PubMed  Google Scholar 

  29. Wise JP, Orenstein JM, Patierno SR: Inhibition of lead chromate clastogenesis by ascorbate: Relationship to particle dissolution and uptake. Carcinogensis 14: 429–434, 1993

    Google Scholar 

  30. Daniel LN, Mao Y, Saffiotti U: Oxidative DNA damage by crystalline silica. Free Radic Biol Med 14: 463–472, 1993

    Article  PubMed  Google Scholar 

  31. Shi X, Dalal NS: Chromium(V) and hydroxyl radical formation during the glutathione reductase-catalyzed reduction of chromium(VI). Biochem Biophys Res Commun 163: 627–634, 1989

    PubMed  Google Scholar 

  32. Shi X, Dalal NS: On the hydroxyl radical formation in the reaction between hydrogen peroxide and biologically generated chromium(V) species. Arch Biochem Biophys 277: 342–350, 1990

    Article  PubMed  Google Scholar 

  33. Shi X, Dalal NS: Evidence for a Fenton-type mechanism for the generation of ·OH radical in the reduction of Cr(VI) in cellular media. Arch Biochem Biophys 281: 90–95, 1990

    PubMed  Google Scholar 

  34. Shi X, Mao Y, Knapton A, Ding M, Rojanasakul Y, Gannett PM, Dalal NS, Liu K: Reaction of Cr(VI) with ascorbate and hydrogen peroxide generates hydroxyl radicals and causes DNA damage: Role of a Cr(IV)-mediated Fenton-like reaction. Carcinogenesis 15: 2475–2478, 1994

    PubMed  Google Scholar 

  35. Buettner GR: ESR parameters of spin adducts. Free Radic Biol Med 3: 259–303, 1987

    PubMed  Google Scholar 

  36. Ione S, Kawanish S: ESR evidence for superoxide, hydroxyl and singlet oxygen produced from hydrogen peroxide and nickel(II) complex of glycylglycyl-L-histidine. Biochem Biophys Res Commun 159: 445–451, 1989

    Article  PubMed  Google Scholar 

  37. Yim MB, Chock PB, Stadtman ER: Copper, zinc superoxide dismutase catalyzes hydroxyl radical production from hydrogen peroxide. Proc Natl Acad Sci USA 87: 5006–5010, 1990

    PubMed  Google Scholar 

  38. Halliwell B: Protection against tissue damage in vivo by deferoxamine: What is its mechanism of activation? Free Radic Biol Med 7: 645–651, 1989

    PubMed  Google Scholar 

  39. Keller RJ, Rush JD, Grover TA: Spectrophotometric and ESR evidence for vanadium(IV) deferoxamine complexes. J Inorg Biochem 41: 268–276, 1991

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Pathology and Physiology Research Branch, Health Effects Laboratory Division, National Institute for Occupational Safety and Health, Morgantown, WV

    Stephen S. Leonard, Val Vallyathan, Vince Castranova & Xianglin Shi

  2. Department of Basic Pharmaceutical Sciences, West Virginia University, Morgantown, WV, USA

    Stephen S. Leonard

Authors
  1. Stephen S. Leonard
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Val Vallyathan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Vince Castranova
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Xianglin Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Leonard, S.S., Vallyathan, V., Castranova, V. et al. Generation of reactive oxygen species in the enzymatic reduction of PbCrO4 and related DNA damage. Mol Cell Biochem 234, 309–315 (2002). https://doi.org/10.1023/A:1015917922980

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1015917922980

Search

Navigation