title: Nasal cytochrome P450 2A: identification, regional localization, and metabolic activity toward hexamethylphosphoramide, a known nasal carcinogen.
abstract: Two members of the cytochrome P450 2A subfamily, CYP2A10 and 2A11, are abundant nasal enzymes previously characterized in rabbit olfactory microsomes. Rabbit CYP2A is active toward a number of nasal toxicants, including the rat nasal procarcinogen hexamethylphosphoramide (HMPA). While P450s immunochemically related to the rabbit CYP2As have been detected in rat and human nasal mucosa, confirmation of these enzymes as members of the CYP2A subfamily and efforts to characterize their ability to bioactivate toxicants have been limited. In the present study, the regional distribution and cell-specific expression of CYP2A in the rat nasal cavity were examined using an antibody to rabbit CYP2A10/11. In sections of the anterior nose, immunoreactive CYP2A was present in ciliated cells of the nasal respiratory epithelium and cuboidal epithelial cells of the nasal transitional epithelium, but was absent in squamous epithelial cells. The most intense immunostaining was observed in the posterior nose. Olfactory sustentacular cells and Bowman's gland cells in sections posterior to the nasal papilla stained most intensely. Western blot analysis revealed that anti-CYP2A10/11 recognized a sharp band of approximately 50 kDa in nasal respiratory and olfactory microsomes, supporting the premise that the antibody is reacting with a cytochrome P450 enzyme. The nasal expression of CYP2A6 mRNA--a member of the human CYP2A subfamily having a high degree of homology to rabbit 2A10 and 2A11--was examined in human surgical patients. Middle turbinectomy tissues--largely composed of nasal respiratory epithelia--from 11 patients were analyzed for the presence of CYP2A6 using reverse transcription-polymerase chain reaction (RT-PCR). Identification of CYP2A6 was confirmed by DNA sequencing of RT-PCR products. CYP2A6 mRNA was detected in all of the human samples analyzed. In additional experiments, human CYP2A6 metabolized HMPA to formaldehyde, suggesting that this compound might cause nasal toxicity in humans. The identification of CYP2A cytochromes in rat and human nasal tissues may have important implications for risk assessment of inhaled xenobiotics.
authors.affiliation: Inhalation Toxicology Research Institute, Albuquerque, New Mexico 87185, USA.
authors.forename: J R
authors.initials: JR
authors.lastname: Thornton-Manning
authors.forename: K J
authors.initials: KJ
authors.lastname: Nikula
authors.forename: J A
authors.initials: JA
authors.lastname: Hotchkiss
authors.forename: K J
authors.initials: KJ
authors.lastname: Avila
authors.forename: K D
authors.initials: KD
authors.lastname: Rohrbacher
authors.forename: X
authors.initials: X
authors.lastname: Ding
authors.forename: A R
authors.initials: AR
authors.lastname: Dahl
mesh_terms: D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases; D001711:Biotransformation; D002273:Carcinogens / Q000493:pharmacokinetics*; D065637:Cytochrome P-450 CYP2A6; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D000072467:Cytochrome P450 Family 2; D006492:Hempa / Q000493:pharmacokinetics*; D006801:Humans; D008168:Lung / Q000378:metabolism; D008297:Male; D008861:Microsomes / Q000378:metabolism; D008862:Microsomes, Liver / Q000378:metabolism; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism*; D009297:Nasal Mucosa / Q000201:enzymology* / Q000648:ultrastructure; D009831:Olfactory Mucosa / Q000201:enzymology / Q000648:ultrastructure; D009928:Organ Specificity; D016133:Polymerase Chain Reaction; D011355:Prodrugs / Q000493:pharmacokinetics; D012333:RNA, Messenger / Q000032:analysis; D011817:Rabbits; D051381:Rats; D011916:Rats, Inbred F344; D013045:Species Specificity
chemical_list: D002273:Carcinogens; D011355:Prodrugs; D012333:RNA, Messenger; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D001189:Aryl Hydrocarbon Hydroxylases; C082663:CYP2A10 protein, Oryctolagus cuniculus; C082665:CYP2A11 protein, Oryctolagus cuniculus; C585710:CYP2A6 protein, human; D065637:Cytochrome P-450 CYP2A6; D000072467:Cytochrome P450 Family 2; D006492:Hempa