N-Acetyltransferases, sulfotransferases and heterocyclic amine activation in the breast.
Heterocyclic amines are mammary carcinogens in rats and their N-hydroxy metabolites are substrates for subsequent metabolic activation by N-acetyltransferases (NAT) and sulfotransferases (SULT) in man. We investigated the expression of these enzymes in human breast tissue and the relationship between NAT genotype and NAT mRNA expression or enzyme activity. Immunohistochemical staining of sections of breast tissue identified expression of NAT1 and NAT2 protein in human mammary epithelial cells, but not in the stroma. We also measured the formation of DNA adducts of the heterocyclic amines 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine in calf thymus DNA after incubation of their promutagenic N-hydroxy metabolites with mammary cytosols prepared from reduction mammoplasty tissue. Experimental observations gained from use of enzyme cofactors and NAT and/or SULT inhibitors on cytosolic enzyme activity, recombinant NAT1 activity and heterocyclic amine-DNA adduct formation suggest that both NAT1 and SULT1A enzymes contribute significantly to the activation of N-hydroxylated heterocyclic amines in mammary tissue. NAT1 mRNA transcript levels were found to be two- to three-fold higher than mRNA transcripts of the NAT2 gene in reduction mammoplasty tissue and mammary epithelial cells. NAT1-specific p-aminobenzoic acid acetylation activity, but not NAT2-specific sulfamethazine acetylation activity, was detectable in mammary cytosols. There was no association apparent between NAT genotype and the levels of NAT mRNA or NAT enzyme activity, or between NAT1 genotype and IQ-DNA adduct formation mediated by mammary cytosols. Western blot analysis of mammary cytosolic protein showed detectable levels of SULT1A1 and SULT1A3.
Williams
J A
JA
Institute of Cancer Research, Haddow Laboratories, Sutton, UK.
Stone
E M
EM
Fakis
G
G
Johnson
N
N
Cordell
J A
JA
Meinl
W
W
Glatt
H
H
Sim
E
E
Phillips
D H
DH
D000588:Amines / Q000493:pharmacokinetics*; D001191:Arylamine N-Acetyltransferase / Q000037:antagonists & inhibitors / Q000235:genetics*; D001711:Biotransformation; D015153:Blotting, Western; D001940:Breast / Q000187:drug effects* / Q000201:enzymology; D002273:Carcinogens / Q000493:pharmacokinetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005838:Genotype; D006801:Humans; D007093:Imidazoles / Q000493:pharmacokinetics; D007150:Immunohistochemistry; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000235:genetics*; D011725:Pyridines / Q000493:pharmacokinetics; D011804:Quinolines / Q000493:pharmacokinetics; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015238:Sulfotransferases / Q000235:genetics*
D000588:Amines; D002273:Carcinogens; D004791:Enzyme Inhibitors; D007093:Imidazoles; D007527:Isoenzymes; D011725:Pyridines; D011804:Quinolines; D012333:RNA, Messenger; C060533:2-hydroxyamino-1-methyl-6-phenylimidazo(4,5-b)pyridine; C029216:2-amino-3-methylimidazo(4,5-f)quinoline; C032012:2-hydroxyamino-3-methylimidazolo(4,5-f)quinoline; D001191:Arylamine N-Acetyltransferase; C089384:N-acetyltransferase 1; C478900:NAT2 protein, human; D015238:Sulfotransferases