Archives of Toxicology

, Volume 93, Issue 7, pp 1893–1902 | Cite as

Bioactivation of the tobacco carcinogens 4-aminobiphenyl (4-ABP) and 2-amino-9H-pyrido[2,3-b]indole (AαC) in human bladder RT4 cells

  • Medjda Bellamri
  • Lihua Yao
  • Radha Bonala
  • Francis Johnson
  • Linda B. Von Weymarn
  • Robert J. TureskyEmail author
Toxicokinetics and Metabolism


Occupational and tobacco exposure to aromatic amines (AAs) including 4-aminobiphenyl (4-ABP) and 2-naphthylamine (2-NA) are associated with bladder cancer (BC) risk. Several epidemiological studies have also reported a possible role for structurally related heterocyclic aromatic amines (HAAs) formed in tobacco smoke or cooked meats with BC risk. We had screened for DNA adducts of 4-ABP, 2-NA, and several prominent HAAs formed in tobacco smoke or grilled meats including 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), 2-amino-3,8-dimethylmidazo[4,5-f]quinoxaline (MeIQx), and 2-amino-9H-pyrido[2,3-b]indole (AαC) in the bladder DNA of BC patients, using liquid chromatography/mass spectrometry. We detected DNA adducts of 4-ABP, but not adducts of the other carcinogens. In this study, we have examined the capacity of RT4 cells, an epithelial human bladder cell line, to bioactivate AAs and HAAs to DNA damaging agents, which may contribute to BC. 4-ABP and AαC formed DNA adducts, but DNA adducts of 2-NA, PhIP, and MeIQx were not detected. 4-ABP DNA adducts were formed at tenfold higher levels than AαC adducts. Pretreatment of RT4 cells with α-naphthoflavone (1–10 µM), a specific cytochrome P450 1 (CYP1) inhibitor, decreased AαC adduct formation by 50% but did not affect the level of 4-ABP adducts. However, cell pretreatment with 8-methoxypsoralen (0.1–1 µM), a potent inhibitor of CYP2A, resulted in a 90% decrease of 4-ABP DNA adducts levels. These data signify that CYP2A and CYP1A isoforms expressed in the target urothelium bioactivate 4-ABP and AαC, respectively, and may be a critical feature of aromatic amine-induced urinary bladder carcinogenesis. The bioactivation of other tobacco and environmental AAs by bladder CYPs and their ensuing bladder DNA damage warrants further study.


Bladder cancer Aromatic amines Heterocyclic aromatic amines 4-Aminobiphenyl 2-Amino-9H-pyrido[2,3-b]indole DNA adduct 



This work is supported by R01ES019564 and R01ES030559 (R. J. T.) from the National Institute of Environmental Health Sciences and by R01CA220367 (R. J. T.) from the National Cancer Institute, National Institutes of Health. Mass spectrometry was carried out in the Analytical Biochemistry Share Resources of the Masonic Cancer Center, the University of Minnesota, funded in part by Cancer Center Support Grant CA-077598. The Turesky laboratory greatfully acknowledges the support of the Masonic Chair in Cancer Causation, University of Minnesota.

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Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

204_2019_2486_MOESM1_ESM.docx (875 kb)
Supplementary material 1 (DOCX 875 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Masonic Cancer Center and Department of Medicinal Chemistry, Cancer and Cardiovascular Research BuildingUniversity of MinnesotaMinneapolisUSA
  2. 2.Department of Pharmacological SciencesStony Brook UniversityStony BrookUSA
  3. 3.Department of ChemistryStony Brook UniversityStony BrookUSA
  4. 4.Department of Biochemistry, Molecular Biology and Biophysics and Masonic Cancer CenterUniversity of MinnesotaMinneapolisUSA

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