Chemical carcinogen induction of DNA-repair synthesis in human peripheral blood monocytes.
Fresh ex vivo cultures of normal human peripheral blood monocytes, which are nonreplicative and known to possess cytochrome P-450 associated mixed-function oxidase activity, were used to assay DNA-excision repair manifested as augmented [3H]thymidine (dThd) incorporation following treatment in culture with diverse mutagenic carcinogens. Untreated monocyte cultures established from pools of 3-6 normal donors incorporated a low level of cytoplasmic [3H]dThd throughout a majority of the cells during an 18-h incubation. This background incorporation into whole cells was 80-90% inhibited by hydroxyurea (HU) at concentrations greater than 5 mM. Dose-related increases in the cumulative 18-h [3H]dThd incorporation in monocytes were observed following treatment with UV, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), methyl methanesulfonate (MMS), mitomycin C (MMC), N-acetoxy-acetylaminofluorene (NA-AAF), aflatoxin B1 (AFB1), benzo[a]pyrene (BaP), and dimethylnitrosamine (DMN). The presence of HU during chemical treatment and throughout this 18 h of incubation with [3H]dThd did not influence the dose-response curves obtained with UV, MMS, NA-AAF and BaP but it increased the input dose of MNNG, MMC, DMN and AFB1 required to give peak repair incorporation. When HU was added to cultures following MNNG damage no interference with repair response was observed. HU apparently influences the extent of DNA damage by direct reactivity with these chemicals or their endogenously generated metabolites rather than inhibiting DNA-repair processes. These results provide evidence that monocytes are enzymatically proficient in base and nucleotide excision pathways and have endogenous capacity to metabolize BaP, AFB1 and DMN to DNA-damaging metabolites. As such, the monocyte is a potentially useful human cell type for detecting genotoxic chemicals and studying individuality in chemical-biological interactions.
D015073:2-Acetylaminofluorene / Q000494:pharmacology; D000348:Aflatoxins / Q000494:pharmacology; D001580:Benzopyrenes / Q000494:pharmacology; D002273:Carcinogens / Q000494:pharmacology*; D002478:Cells, Cultured; D004260:DNA Repair / Q000187:drug effects*; D004261:DNA Replication / Q000187:drug effects*; D004128:Dimethylnitrosamine / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008741:Methyl Methanesulfonate / Q000494:pharmacology; D008769:Methylnitronitrosoguanidine / Q000494:pharmacology; D008937:Mitomycins / Q000494:pharmacology; D009000:Monocytes / Q000378:metabolism*; D009153:Mutagens
D000348:Aflatoxins; D001580:Benzopyrenes; D002273:Carcinogens; D008937:Mitomycins; D009153:Mutagens; D008769:Methylnitronitrosoguanidine; D015073:2-Acetylaminofluorene; D008741:Methyl Methanesulfonate; D004128:Dimethylnitrosamine