Abstract
4,4′-Methylenebis(2-chloroaniline) or MOCA is an aromatic amine used primarily in polyurethane and rubber industry. MOCA has been linked to hepatomas in animal studies while limited epidemiologic studies reported the association of exposure to MOCA and urinary bladder and breast cancer. We investigated MOCA-induced genotoxicity and oxidative stress in DNA repair-deficient Chinese hamster ovary (CHO) cells stably transfected with human metabolizing enzymes CYP1A2 and N-acetyltransferase 2 (NAT2) variants as well as in rapid, intermediate, and slow NAT2 acetylator cryopreserved human hepatocytes. N-acetylation of MOCA was highest in UV5/1A2/NAT2*4 followed by UV5/1A2/NAT2*7B and UV5/1A2/NAT2*5B CHO cells. Human hepatocytes showed a NAT2 genotype-dependent response with highest N-acetylation in rapid acetylators followed by intermediate and slow acetylators. MOCA induced higher levels of mutagenesis and DNA damage in UV5/1A2/NAT2*7B compared to UV5/1A2/NAT2*4 and UV5/1A2/NAT2*5B cells (p < 0.0001). MOCA also induced higher levels of oxidative stress in UV5/1A2/NAT2*7B cells. MOCA caused concentration-dependent increase in DNA damage in cryopreserved human hepatocytes (linear trend p < 0.001) which was NAT2 genotype dependent i.e., highest in rapid acetylators, lower in intermediate acetylators, and lowest in slow acetylators (p < 0.0001). Our findings show that N-acetylation and genotoxicity of MOCA is NAT2 genotype dependent and suggest that individuals possessing NAT2*7B are at higher risk to MOCA-induced mutagenicity. DNA damage, and oxidative stress. They confirm significant differences in genotoxicity between the NAT2*5B and NAT2*7B alleles, both of which are associated with slow acetylator phenotype.
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Data availability
The datasets for this study are available from the corresponding author upon request.
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Acknowledgements
This work was partially supported by United States Public Health Service Grants P20-GM113226, P30-ES030283 and P42-ES023716. Portions of this work constituted partial fulfilment of the PhD in pharmacology and toxicology by Mariam Habil at the University of Louisville.
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MRH: writing the original draft, conducted experiments, data visualization formal analysis, review, and editing. RASG: conducted experiments, data visualization, formal analysis investigation, writing, review, and editing. MAD: conducted experiments, formal analysis, investigation, writing, review, and editing. DWH: conceptualization, methodology, validation, formal analysis, resources, writing, review and editing, visualization, supervision.
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Habil, M.R., Salazar-González, R.A., Doll, M.A. et al. Effect of N-acetyltransferase 2 genetic polymorphism on 4,4′-methylenebis(2-chloroaniline)-induced genotoxicity and oxidative stress. Arch Toxicol 97, 1773–1781 (2023). https://doi.org/10.1007/s00204-023-03508-1
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DOI: https://doi.org/10.1007/s00204-023-03508-1