Archives of Toxicology

, Volume 70, Issue 3–4, pp 135–144 | Cite as

Effects of benzene metabolite treatment on granulocytic differentiation and DNA adduct formation in HL-60 cells

  • C. C. Hedli
  • N. R. Rao
  • K. R. Reuhl
  • C. M. Witmer
  • R. Snyder
Original Investigation

Abstract

Reactive metabolites of benzene (BZ) play important roles in BZ-induced hematotoxicity. Although reactive metabolites of BZ covalently bind to DNA, the significance of DNA adduct formation in the mechanism of BZ toxicity is not clear. These studies investigated the covalent binding of the BZ metabolites hydroquinone (HQ) and 1,2,4-benzenetriol(BT) using the DNA [32P]postlabeling method and explored the potential relationship between DNA adduct formation and cell differentiation in human promyelocytic leukemia (HL-60) cells, a model system for studying hematopoiesis. Maturation of HL-60 cells to granulocytes, as assessed by light and electron microscopy, was significantly inhibited in cells that were pretreated with HQ or BT prior to inducing differentiation with retionic acid (RA). The capacity of RA-induced cells to phagocytose sheep red blood cells (RBC) and to reduce nitroblue tetrazolium (NBT), two functional parameters characteristic of mature, differentiated neutrophils, was also inhibited in cells pretreated with HQ or BT. These BZ metabolite treatments induced DNA adduct formation in HQ- but not in BT-treated cells. These results indicate that whereas HQ and BT each block granulocytic differentiation in HL-60 cells, DNA adducts were observed only following HQ treatment. Thus DNA adduct formation may be important in HQ but not in BT toxicity.

Key words

Benzene Metabolite Granulocytic differentiation DNA adducts HL-60 cells 

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References

  1. Bodell WJ, Levay G, Pongracz K (1993) Investigation of Benzene-DNA adducts and their detection in human bone marrow. Envir Health Perspect 99: 241–244CrossRefGoogle Scholar
  2. Breitman TR, Selonic SE, Collins SJ (1980) Induction of differentiation of the human promyelocytic leukemia cell line (HL-60) by retinoic acid. Proc Natl Acad Sci USA 77: 2936–2940PubMedCrossRefGoogle Scholar
  3. Collins SJ (1987) The HL-60 cell line: proliferation, differentiation, and cellular oncogene expression. Blood 70: 1233–1244PubMedGoogle Scholar
  4. Davis L, Kuehl M, Battey J (1994) Preparation of genomic DNA from tissue culture cells. In: Davis L, Kuehl M, Battey J (eds) Basic methods in molecular biology, 2nd Edition. Appleton and Lange, East Norwalk, Conneticut, pp 310–313Google Scholar
  5. Forni A, Pacifico E, Limonata A (1971a) Chromosome studies in workers exposed to benzene or toluene or both. Arch Environ Health 22: 373–378PubMedGoogle Scholar
  6. Forni AM, Cappellini A, Pacifico E, Vigliani EC (1971b) Chromosome changes and their evolution in subjects with past exposures to benzene. Arch Environ Health 23: 385–391PubMedGoogle Scholar
  7. Gowda SD, Koler RD, Bagby GC (1986) Regulation of c-myc expression during growth and differentiation of normal and leukemic human myeloid progenitor cells. J Clin Invest 77: 271–278PubMedCrossRefGoogle Scholar
  8. Gupta RC (1985) Enhanced sensitivity of 32P-postlabeling analysis of aromatic carcinogen-DNA adducts. Cancer Res 5: 5656–5662Google Scholar
  9. Gupta RC, Reddy MV, Randerath K (1982) 32P-postlabeling analysis of non-radioactive aromatic carcinogen-DNA adducts. Carcinogenesis 3: 1081–1092PubMedCrossRefGoogle Scholar
  10. Johnson RA, Walseth TF (1979) The enzymatic preparation of [α-32P]ATP, [α-32P]GTP, [32P]CAMP, [32P]cGMP, and their use in assay of adenylate and guanylate cyclases, and cyclic nucleotide phosphodiesterases. Adv Cyc Nucl Res 117: 271–279Google Scholar
  11. Jowa L, Witz G, Snyder R, Winkle S, Kalf GF (1990) Synthesis and characterization of deoxyguanosine-benzoquinone adducts. J Appl Toxicol: 10: 47–54PubMedCrossRefGoogle Scholar
  12. Kalf GF, O’Connor A (1993) The effects of benzene and hydroquinone on myeloid differentiation in HL-60 promyelocytic leukemia cells. Leuk Lymph 11: 331–338CrossRefGoogle Scholar
  13. Kolanchana P, Subrahamanyam VV, Meyer KB, Zhang L, Smith MT (1993) Benzene and its phenolic metabolites produce oxidative damage in HL-60 cells in vitro and in the bone marrow in vivo. Cancer Res 53: 1023–1026Google Scholar
  14. LeBeau M, Larson RA (1991) Cytogenetics and neoplasia. In: Hoffman R, Benz EJ, Shattil SJ, Furie B, Cohen HJ (eds) Hematology basic principles and practice, vol. 50. Churchill Livingstone, New York, pp 638–655Google Scholar
  15. Levay G, Bodell WJ (1992) Potentiation of DNA adduct formation in HL-60 cells by combinations of benzene metabolites. Proc Natl Acad Sci USA 89: 7105–7109PubMedCrossRefGoogle Scholar
  16. Levay G, Pongracz K, Bodell WJ (1991) Detection of DNA adducts in HL-60 cells treated with hydroquinone and p-benzoquinone by 32P-postlabeling. Carcinogenesis 12: 1181–1186PubMedCrossRefGoogle Scholar
  17. Levay G, Ross D, Bodell WJ (1993) Peroxidase activation of hydroquinone results in the formation of DNA adducts in Hl-60 cells, mouse bone marrow macrophages and human bone marrow. Carcinogenesis 14: 2329–2334PubMedCrossRefGoogle Scholar
  18. Lutz WK, Schlatter CH (1977) Mechanism of the carcinogenic action of benzene; irreversible binding to rat liver DNA. Chem-Biol Interact 18: 241–245PubMedCrossRefGoogle Scholar
  19. McBride TJ, Preston BD, Loeb LA (1991) Mutagenic spectrum resulting from DNA damage by oxygen radicals. Biochemistry 30: 207–213PubMedCrossRefGoogle Scholar
  20. Oliveira NL, Kalf GF (1992) The induced differentiation of HL-60 cells to monocyte/macrophages is inhibited by hydroquinone, a hematotoxic metabolite of benzene. Blood 79: 627–633PubMedGoogle Scholar
  21. Pellack-Walker P, Walker JK, Evans HH, Blumer JL (1985) Relationship between the oxidation potential of benzene metabolites and their inhibitory effect on DNA synthesis in L5178YS cells. Mol Pharmacol 28: 560–566PubMedGoogle Scholar
  22. Pollini G, Collombi R (1964) Chromosomal damage in lymphocytes during benzene hemopathy. Med J Lav 55: 641–654Google Scholar
  23. Rao NR, Snyder R (1995) Oxidative modifications, produced in HL-60 cells upon exposure to benzene metabolites. J Appl Toxicol 15: 403–409PubMedCrossRefGoogle Scholar
  24. Reddy MV, Randerath K (1986). Nuclease P1-mediated enhancement of sensitivity of 32P-postlabeling test for structurally diverse DNA adducts. Carcinogenesis 7: 1543–1551PubMedCrossRefGoogle Scholar
  25. Reddy MV, Blackburn GR, Irwin SE, Kommineni C, Mackerer CR, Mehlman MA (1989) A method for in vitro culture of rat Zymbal gland: use in mechanistic studies of benzene carcinogenesis in combination with 32P-postlabeling. Environ Health Perspect 82: 239–248PubMedCrossRefGoogle Scholar
  26. Rickert DE, Baker TS, Bus JS, Barrow CS, Irons RD (1979) Benzene disposition in the rat after exposure by inhalation. Toxicol Appl Pharmacol 49: 417–423PubMedCrossRefGoogle Scholar
  27. Rushmore T, Kalf G, Snyder R (1984) Covalent binding of benzene and its metabolites to DNA in rabbit bone marrow mitochondria in vitro. Chem.-Biol Interact 49: 133–154PubMedCrossRefGoogle Scholar
  28. Sasiakek M (1992) Nonrandom distribution of breakpoints in the karyotypes of workers occupationally exposed to benzene. Environ Health Perspect 97: 255–257CrossRefGoogle Scholar
  29. Sato T (1968) Modified method for lead staining of thin sections. J Electron Microsc 17: 158–159Google Scholar
  30. Snyder R, Lee EW, Kocsis JJ (1978) Binding of labeled benzene metabolites to mouse liver and bone marrow. Res Commun Chem Pathol Pharmacol 20: 191–194PubMedGoogle Scholar
  31. Snyder R, Witz G, Goldstein BD (1993) The toxicology of benzene. Environ Health Perspect 100: 293–306PubMedCrossRefGoogle Scholar
  32. Teebor GW, Boorstein RJ, Cadet J (1988) The repairability of oxidative free radical mediated damage to DNA: a review. Int J Rad Biol 54: 131–150PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • C. C. Hedli
    • 1
  • N. R. Rao
    • 2
  • K. R. Reuhl
    • 1
  • C. M. Witmer
    • 1
  • R. Snyder
    • 1
  1. 1.Joint Graduate Program in ToxicologyRutgers University, UMDNJ-RWJMSPiscatawayUSA
  2. 2.Department of PharmacologyNASA/Johnson Space CenterHoustonUSA

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