A comparison of the HL-60 cell line and human granulocytes to effect the bioactivation of arylamines and related xenobiotics. The binding of 2-aminofluorene to nucleic acids as the result of the respiratory burst.
Studies were made on the ability of the leukemic cell line, HL-60, to substitute for normal human granulocytes in research concerned with the bioactivation of arylamines. The arylamine carcinogen, 2-aminofluorene (2-AF), was used as the model substrate in the form of 2-[9-14C]AF, and was incubated with HL-60 cell cultures, both in the presence and absence of phorbol myristate acetate (PMA) which induces the respiratory burst. The HL-60 cultures were generally employed after having been induced to undergo differentiation to neutrophils by the action of dimethyl sulfoxide (DMSO). Comparisons of the amounts of DNA and RNA binding by 2-AF between HL-60 and normal human granulocyte cultures demonstrated close similarities in the amount and nature of nucleic acid binding by this arylamine substrate. HL-60 cells that had been induced to differentiate to neutrophils to the extent of about 80% showed high levels of the respiratory burst along with extensive covalent binding of 2-[9-14C]AF to cellular nucleic acids. Although normal human granulocytes tended to metabolize 2-AF slightly faster than did highly differentiated HL-60 cells, the extent of nucleic acid binding relative to the amount of 2-AF metabolized was similar. A major difference in the metabolic fate of 2-AF in these cell cultures was the unique ability of HL-60 cultures at all stages of differentiation to effect the slow N-acetylation of 2-AF to give 2-acetylaminofluorene (2-AAF). Extensive analyses of incubation extracts showed that the major differences in apparent metabolites were quantitative. With few exceptions, both activated HL-60 and granulocyte cell cultures produced the same metabolites, most of which remain unidentified. Studies with inhibitors such as catalase, superoxide dismutase and azide ion further suggest that these two related cell cultures metabolize 2-AF in similar manner. The DMSO-differentiated HL-60 culture is proposed as a convenient model with which to investigate the metabolism and bioactivation of arylamines by human granulocytes or pure neutrophils.
Corbett
M D
MD
Food Science and Human Nutrition Department, University of Florida, Gainesville 32611.
Hannothiaux
M H
MH
Corbett
B R
BR
Quintana
S J
SJ
D000588:Amines / Q000097:blood / Q000378:metabolism*; D002273:Carcinogens / Q000378:metabolism*; D002454:Cell Differentiation; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004247:DNA / Q000378:metabolism*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005449:Fluorenes / Q000378:metabolism*; D006098:Granulocytes / Q000378:metabolism*; D006801:Humans; D007942:Leukemia, Experimental / Q000378:metabolism* / Q000473:pathology; D007951:Leukemia, Myeloid / Q000378:metabolism* / Q000473:pathology; D009504:Neutrophils / Q000166:cytology / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D010101:Oxygen Consumption; D011485:Protein Binding; D012313:RNA / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D015262:Xenobiotics / Q000097:blood / Q000378:metabolism*
D000588:Amines; D002273:Carcinogens; D005449:Fluorenes; D015262:Xenobiotics; C012177:2-aminofluorene; D012313:RNA; D004247:DNA; D013755:Tetradecanoylphorbol Acetate; D010100:Oxygen; D004121:Dimethyl Sulfoxide