Human cytochromes P450 1A1 and 1B1 catalyze ring oxidation but not nitroreduction of environmental pollutant mononitropyrene isomers in primary cultures of human breast cells and cultured MCF-10A and MCF-7 cell lines.
The position of the nitro group determines the relative carcinogenic activities of mono-nitropyrene isomers (mono-NPs) in the rat mammary gland. To determine whether the results obtained in rodents treated with these environmental pollutants can be applicable to humans, we examined their metabolic activation in primary cultures of human breast cells derived from reduction mammoplasty, as well as in the cultured human breast cancer cell line MCF-7 and the immortalized human mammary epithelial cell line MCF-10A. Primary cultures as well as cell lines were competent in metabolizing all three isomers via both ring oxidation and nitro reduction pathways. Qualitatively similar metabolic patterns were observed but quantitative differences were evident. On the basis of cochromatography with synthetic standards in two HPLC systems, metabolites of 1-NP were identified as 1-OH-Py, 3-, 6-, and 8-OH-1-NP and 1-AP. In the case of 2-NP, 6-OH-2-NP and 2-AP were identified. 4-NP was metabolized to 9,10-DHD-4-NP, Py-4,5-Q, 9,10-Q-4-NP, 9/10-OH-4-NP, 6/ 8-OH-4-NP, and 4-AP. Varying degrees of sulfate and glucuronide conjugation of mono-NP metabolites were detected. In MCF-7 cells, we found that 1-, 2-, and 4-NP bind to DNA at levels of 68, 17, and 132 pmol/mg DNA, respectively. Following HPLC analysis of the DNA hydrolysates, we detected multiple DNA adducts including those derived from nitro reduction of 2- and 4-NP; however, none was detected in the case of 1-NP. To determine the P450 enzymes responsible for the metabolic activation of these carcinogens, we incubated [(3)H]mono-NPs with recombinant human P450 1A1 or 1B1. Metabolites identified were primarily derived from ring oxidation; both P450s 1A1 and 1B1 yielded similar metabolic profiles. This is the first report demonstrating that human breast (target organ) cells, immortalized human mammary epithelial cell line MCF-10A, and breast cancer cell line MCF-7 are capable of activating mono-NPs to metabolites that can damage DNA.
Division of Cancer Etiology and Prevention, Institute for Cancer Prevention, Valhalla, New York 10595, USA.
Yuan-Wan
YW
Sun
F Peter
FP
Guengerich
Arun K
AK
Sharma
Telih
T
Boyiri
Shantu
S
Amin
Karam
K
el-Bayoumy
D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics / Q000378:metabolism*; D001940:Breast / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D001943:Breast Neoplasms / Q000201:enzymology*; D045744:Cell Line, Tumor; D019363:Cytochrome P-450 CYP1A1 / Q000235:genetics / Q000378:metabolism*; D065633:Cytochrome P-450 CYP1B1; D018736:DNA Adducts / Q000378:metabolism; D004785:Environmental Pollutants / Q000378:metabolism*; D004847:Epithelial Cells / Q000201:enzymology; D005260:Female; D006801:Humans; D007536:Isomerism; D009574:Nitro Compounds / Q000378:metabolism; D010084:Oxidation-Reduction; D011721:Pyrenes / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014407:Tumor Cells, Cultured
D018736:DNA Adducts; D004785:Environmental Pollutants; D009574:Nitro Compounds; D011721:Pyrenes; D011994:Recombinant Proteins; D001189:Aryl Hydrocarbon Hydroxylases; C585496:CYP1B1 protein, human; C585499:Cyp1b1 protein, rat; D019363:Cytochrome P-450 CYP1A1; D065633:Cytochrome P-450 CYP1B1