title: CYP2A13 in human respiratory tissues and lung cancers: an immunohistochemical study with a new peptide-specific antibody.
abstract: Human cytochrome P450 2A13 (CYP2A13) is highly efficient in the metabolic activation of a tobacco-specific carcinogen, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), and another potent carcinogen, aflatoxin B1 (AFB1). Although previous studies demonstrated that CYP2A13 mRNA is predominantly expressed in human respiratory tissues, expression of CYP2A13 protein in these tissues and the involved cell types have not been determined because of the lack of CYP2A13-specific antibodies. To explore the toxicological and physiological function of CYP2A13, it is important to understand the tissue/cellular distribution of CYP2A13 protein. In this study, we generated a peptide-specific antibody against human CYP2A13 and demonstrated by immunoblot analysis that this antibody does not cross-react with heterologously expressed human CYP2A6 and mouse CYP2A5 proteins, both sharing a high degree of amino acid sequence similarity with CYP2A13. Nor does the antibody cross-react with heterologously expressed human CYP3A4, CYP2S1, or any of the cytochrome P450 enzymes present in the human liver microsomes. Using this highly specific antibody for immunohistochemical staining, we detected a high level of CYP2A13 protein expression in the epithelial cells of human bronchus and trachea, but a rare distribution in the alveolar cells. There was little expression of CYP2A13 protein in different types of lung cancers. In consideration of the high efficiency of CYP2A13 in NNK metabolic activation, our result is consistent with the reported observations that most smoking-related human lung cancers are bronchogenic and supports that CYP2A13-catalyzed in situ activation may play a critical role in human lung carcinogenesis related to NNK and AFB1 exposure.
authors.affiliation: School of Public Health, Environmental and Occupational Health Sciences Institute, University of Medicine and Dentistry of New Jersey, Room 385, 683 Hoes Lane West, Piscataway, NJ 08854, USA.
authors.forename: Liang-Ru
authors.initials: LR
authors.lastname: Zhu
authors.forename: Paul E
authors.initials: PE
authors.lastname: Thomas
authors.forename: Gang
authors.initials: G
authors.lastname: Lu
authors.forename: Kenneth R
authors.initials: KR
authors.lastname: Reuhl
authors.forename: Guang-Yu
authors.initials: GY
authors.lastname: Yang
authors.forename: Li-Dong
authors.initials: LD
authors.lastname: Wang
authors.forename: Shou-Lin
authors.initials: SL
authors.lastname: Wang
authors.forename: Chung S
authors.initials: CS
authors.lastname: Yang
authors.forename: Xiao-Yang
authors.initials: XY
authors.lastname: He
authors.forename: Jun-Yan
authors.initials: JY
authors.lastname: Hong
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D001189:Aryl Hydrocarbon Hydroxylases / Q000276:immunology / Q000378:metabolism*; D001980:Bronchi / Q000201:enzymology*; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D007150:Immunohistochemistry / Q000379:methods; D007527:Isoenzymes / Q000276:immunology / Q000378:metabolism; D008175:Lung Neoplasms / Q000201:enzymology* / Q000473:pathology; D051379:Mice; D008862:Microsomes, Liver / Q000201:enzymology; D010455:Peptides / Q000276:immunology; D011650:Pulmonary Alveoli / Q000201:enzymology*; D014132:Trachea / Q000201:enzymology*
chemical_list: D000906:Antibodies; D007527:Isoenzymes; D010455:Peptides; D001189:Aryl Hydrocarbon Hydroxylases; C414206:CYP2A13 protein, human