Effects of benzene metabolite treatment on granulocytic differentiation and DNA adduct formation in HL-60 cells
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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 wordsBenzene Metabolite Granulocytic differentiation DNA adducts HL-60 cells
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- 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
- Gupta RC (1985) Enhanced sensitivity of 32P-postlabeling analysis of aromatic carcinogen-DNA adducts. Cancer Res 5: 5656–5662Google Scholar
- 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
- 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
- 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
- Pollini G, Collombi R (1964) Chromosomal damage in lymphocytes during benzene hemopathy. Med J Lav 55: 641–654Google Scholar
- Sato T (1968) Modified method for lead staining of thin sections. J Electron Microsc 17: 158–159Google Scholar