{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,2]],"date-time":"2026-01-02T19:37:52Z","timestamp":1767382672876,"version":"build-2065373602"},"reference-count":43,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2017,10,13]],"date-time":"2017-10-13T00:00:00Z","timestamp":1507852800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81372956","81573194"],"award-info":[{"award-number":["81372956","81573194"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Natural Science Foundation of Jiangsu Universities","award":["14KJA330002"],"award-info":[{"award-number":["14KJA330002"]}]},{"DOI":"10.13039\/501100004608","name":"Natural Science Foundation of Jiangsu province","doi-asserted-by":"publisher","award":["BK20151555"],"award-info":[{"award-number":["BK20151555"]}],"id":[{"id":"10.13039\/501100004608","id-type":"DOI","asserted-by":"publisher"}]},{"name":"333 Advance Talents Projects of Jiangsu Province","award":["BRA2014285"],"award-info":[{"award-number":["BRA2014285"]}]},{"name":"Six talents peak project of Jiangsu province","award":["DG216D5047"],"award-info":[{"award-number":["DG216D5047"]}]},{"name":"Top-notch Academic Programs Project of Jiangsu Higher Education Institutions (TAPP)","award":["PPZY2015A067"],"award-info":[{"award-number":["PPZY2015A067"]}]},{"name":"a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD), China"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJERPH"],"abstract":"<jats:p>Cytochrome P450 2A13 (CYP2A13), an extrahepatic enzyme mainly expressed in the human respiratory system, has been reported to mediate the metabolism and toxicity of cigarette smoke. We previously found that nicotine inhibited 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) metabolism by CYP2A13, but its influence on other components of cigarette smoke remains unclear. The nicotine component of cigarette smoke extract (CSE) was separated, purified, and identified using high-performance liquid chromatography (HPLC) and ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS\/MS), splitting CSE into a nicotine section (CSE-N) and nicotine-free section (CSE-O). Cell viability and apoptosis by Cell Counting Kit-8 (CCK-8) and flow cytometry assays were conducted on immortalized human bronchial epithelial (BEAS-2B) cells stably expressing CYP2A13 (B-2A13) or vector (B-V), respectively. Interestingly, CSE and CSE-O were toxic to BEAS-2B cells whereas CSE-N showed less cytotoxicity. CSE-O was more toxic to B-2A13 cells than to B-V cells (IC50 of 2.49% vs. 7.06%), which was flatted by 8-methoxypsoralen (8-MOP), a CYP inhibitor. CSE-O rather than CSE or CSE-N increased apoptosis of B-2A13 cells rather than B-V cells. Accordingly, compared to CSE-N and CSE, CSE-O significantly changed the expression of three pairs of pro- and anti-apoptotic proteins, Bcl-2 Associated X Protein\/B cell lymphoma-2 (Bax\/Bcl-2), Cleaved Poly (Adenosine Diphosphate-Ribose) Polymerase\/Poly (Adenosine Diphosphate-Ribose) Polymerase (C-PARP\/PARP), and C-caspase-3\/caspase-3, in B-2A13 cells. In addition, recombination of CSE-N and CSE-O (CSE-O\/N) showed similar cytotoxicity and apoptosis to the original CSE. These results demonstrate that the nicotine component decreases the metabolic activation of CYP2A13 to CSE and aids in understanding the critical role of CYP2A13 in human respiratory diseases caused by cigarette smoking.<\/jats:p>","DOI":"10.3390\/ijerph14101221","type":"journal-article","created":{"date-parts":[[2017,10,13]],"date-time":"2017-10-13T11:34:09Z","timestamp":1507894449000},"page":"1221","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":26,"title":["Nicotine Component of Cigarette Smoke Extract (CSE) Decreases the Cytotoxicity of CSE in BEAS-2B Cells Stably Expressing Human Cytochrome P450 2A13"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3399-9933","authenticated-orcid":false,"given":"Minghui","family":"Ji","sequence":"first","affiliation":[{"name":"Key Lab of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, 101 Longmian Avenue, Nanjing 211166, China"},{"name":"State Key Lab of Reproductive Medicine, Institute of Toxicology, Nanjing Medical University, 101 Longmian Avenue, Nanjing 211166, China"},{"name":"School of Nursing, Nanjing Medical University, 101 Longmian Avenue, Nanjing 211166, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yudong","family":"Zhang","sequence":"additional","affiliation":[{"name":"Key Lab of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, 101 Longmian Avenue, Nanjing 211166, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8324-7310","authenticated-orcid":false,"given":"Na","family":"Li","sequence":"additional","affiliation":[{"name":"Key Lab of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, 101 Longmian Avenue, Nanjing 211166, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chao","family":"Wang","sequence":"additional","affiliation":[{"name":"Key Lab of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, 101 Longmian Avenue, Nanjing 211166, China"},{"name":"State Key Lab of Reproductive Medicine, Institute of Toxicology, Nanjing Medical University, 101 Longmian Avenue, Nanjing 211166, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rong","family":"Xia","sequence":"additional","affiliation":[{"name":"Key Lab of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, 101 Longmian Avenue, Nanjing 211166, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9685-5753","authenticated-orcid":false,"given":"Zhan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Key Lab of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, 101 Longmian Avenue, Nanjing 211166, China"},{"name":"State Key Lab of Reproductive Medicine, Institute of Toxicology, Nanjing Medical University, 101 Longmian Avenue, Nanjing 211166, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shou-Lin","family":"Wang","sequence":"additional","affiliation":[{"name":"Key Lab of Modern Toxicology of Ministry of Education, School of Public Health, Nanjing Medical University, 101 Longmian Avenue, Nanjing 211166, China"},{"name":"State Key Lab of Reproductive Medicine, Institute of Toxicology, Nanjing Medical University, 101 Longmian Avenue, Nanjing 211166, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2017,10,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"966","DOI":"10.1016\/S0140-6736(15)60264-1","article-title":"Global trends and projections for tobacco use, 1990\u20132025: An analysis of smoking indicators from the WHO Comprehensive Information Systems for Tobacco Control","volume":"385","author":"Bilano","year":"2015","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1447","DOI":"10.1016\/S0140-6736(15)00340-2","article-title":"Contrasting male and female trends in tobacco-attributed mortality in China: Evidence from successive nationwide prospective cohort studies","volume":"386","author":"Chen","year":"2015","journal-title":"Lancet"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1422","DOI":"10.1016\/S0140-6736(15)00416-X","article-title":"Smoking cessation for Chinese men and prevention for women","volume":"386","author":"Koplan","year":"2015","journal-title":"Lancet"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2123","DOI":"10.1016\/S0140-6736(15)60996-5","article-title":"Tobacco in China: Taming the smoking dragon","volume":"385","author":"Schwartlander","year":"2015","journal-title":"Lancet"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"10398","DOI":"10.1039\/C5NR01496D","article-title":"Silver nanowire interactions with primary human alveolar type-II epithelial cell secretions: Contrasting bioreactivity with human alveolar type-I and type-II epithelial cells","volume":"7","author":"Sweeney","year":"2015","journal-title":"Nanoscale"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"613","DOI":"10.3390\/ijerph8020613","article-title":"Hazardous compounds in tobacco smoke","volume":"8","author":"Talhout","year":"2011","journal-title":"Int. J. Environ. Res. Public Health"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"64","DOI":"10.3390\/ijerph120100064","article-title":"Nnk, a tobacco-specific carcinogen, inhibits the expression of lysyl oxidase, a tumor suppressor","volume":"12","author":"Cheng","year":"2014","journal-title":"Int. J. Environ. Res. Public Health"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Leon, M.E., Peruga, A., McNeill, A., Kralikova, E., Guha, N., Minozzi, S., Espina, C., and Schuz, J. (2015). European Code against Cancer, 4th edition: Tobacco and cancer. Cancer Epidemiol., S20\u2013S33.","DOI":"10.1016\/j.canep.2015.06.001"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1046\/j.1440-1843.2003.00453.x","article-title":"Respiratory health effects of exposure to environmental tobacco smoke","volume":"8","year":"2003","journal-title":"Respirology"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2584","DOI":"10.1093\/carcin\/bgu182","article-title":"Identification of cytochrome P450 enzymes critical for lung tumorigenesis by the tobacco-specific carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK): Insights from a novel Cyp2abfgs-null mouse","volume":"35","author":"Li","year":"2014","journal-title":"Carcinogenesis"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1539","DOI":"10.2741\/s350","article-title":"Cytochrome P450 in non-small cell lung cancer related to exogenous chemical metabolism","volume":"4","author":"Oyama","year":"2012","journal-title":"Front. Biosci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1672","DOI":"10.1124\/dmd.106.011049","article-title":"Cyp2a13 in human respiratory tissues and lung cancers: An immunohistochemical study with a new peptide-specific antibody","volume":"34","author":"Zhu","year":"2006","journal-title":"Drug Metab. Dispos."},{"key":"ref_13","first-page":"8057","article-title":"Substantial reduction in risk of lung adenocarcinoma associated with genetic polymorphism in Cyp2A13, the most active cytochrome P450 for the metabolic activation of tobacco-specific carcinogen NNK","volume":"63","author":"Wang","year":"2003","journal-title":"Cancer Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2316","DOI":"10.1124\/dmd.108.022822","article-title":"Characterization of Cyp2A13*2, a variant cytochrome P450 allele previously found to be associated with decreased incidences of lung adenocarcinoma in smokers","volume":"36","author":"Zhang","year":"2008","journal-title":"Drug Metab. Dispos."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1986","DOI":"10.1096\/fj.14-264580","article-title":"Pulmonary expression of Cyp2A13 and ABCB1 is regulated by FOXA2, and their genetic interaction is associated with lung cancer","volume":"29","author":"Xiang","year":"2015","journal-title":"FASEB J."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1093\/carcin\/bgt269","article-title":"Role of Cyp2A13 in the bioactivation and lung tumorigenicity of the tobacco-specific lung procarcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone: In vivo studies using a Cyp2A13-humanized mouse model","volume":"35","author":"Megaraj","year":"2014","journal-title":"Carcinogenesis"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"95","DOI":"10.1021\/tx049847p","article-title":"Cytochrome P450 enzymes as catalysts of metabolism of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone, a tobacco specific carcinogen","volume":"18","author":"Jalas","year":"2005","journal-title":"Chem. Res. Toxicol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"211","DOI":"10.3109\/00498254.2015.1069419","article-title":"Oxidation of pyrene, 1-hydroxypyrene, 1-nitropyrene and 1-acetylpyrene by human cytochrome P450 2A13","volume":"46","author":"Shimada","year":"2016","journal-title":"Xenobiotica"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"921","DOI":"10.1093\/carcin\/5.7.921","article-title":"Carcinogenicity in mice of a mutagenic compound, 2-amino-3-methylimidazo[4,5-f]quinoline, from broiled sardine, cooked beef and beef extract","volume":"5","author":"Ohgaki","year":"1984","journal-title":"Carcinogenesis"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Benowitz, N.L., Hukkanen, J., and Jacob, P. (2009). Nicotine Chemistry, Metabolism, Kinetics and Biomarkers. Handb. Exp. Pharmacol., 29\u201360.","DOI":"10.1007\/978-3-540-69248-5_2"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1166","DOI":"10.1124\/dmd.105.004549","article-title":"Nicotine 5\u2019-oxidation and methyl oxidation by P450 2A enzymes","volume":"33","author":"Murphy","year":"2005","journal-title":"Drug Metab. Dispos."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1016\/j.tox.2012.06.010","article-title":"Cytochrome P450 2A13 mediates aflatoxin B1-induced cytotoxicity and apoptosis in human bronchial epithelial cells","volume":"300","author":"Yang","year":"2012","journal-title":"Toxicology"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1124\/jpet.105.091306","article-title":"Inactivation of CYP2A6 and CYP2A13 during Nicotine Metabolism","volume":"316","author":"Brown","year":"2006","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Comer, D.M., Elborn, J.S., and Ennis, M. (2014). Inflammatory and cytotoxic effects of acrolein, nicotine, acetylaldehyde and cigarette smoke extract on human nasal epithelial cells. BMC Pulm. Med., 14.","DOI":"10.1186\/1471-2466-14-32"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1007\/s11481-011-9283-6","article-title":"A LC-MS\/MS Method for Concurrent Determination of Nicotine Metabolites and Role of CYP2A6 in Nicotine Metabolism in U937 Macrophages: Implications in Oxidative Stress in HIV + Smokers","volume":"7","author":"Jin","year":"2012","journal-title":"J. Neuroimmune Pharmacol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1697","DOI":"10.1007\/s00204-013-1108-3","article-title":"Cytochrome P450 2A13 is an efficient enzyme in metabolic activation of aflatoxin G1 in human bronchial epithelial cells","volume":"87","author":"Zhang","year":"2013","journal-title":"Arch. Toxicol."},{"key":"ref_27","first-page":"621","article-title":"Effects of 8-methoxypsoralen on cytochrome P450 2A13","volume":"26","author":"Zhang","year":"2005","journal-title":"Carcinogenesis"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.tox.2013.08.015","article-title":"Identification of stable cytotoxic factors in the gas phase extract of cigarette smoke and pharmacological characterization of their cytotoxicity","volume":"314","author":"Noya","year":"2013","journal-title":"Toxicology"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1254\/jphs.11166FP","article-title":"Nicotine- and tar-free cigarette smoke induces cell damage through reactive oxygen species newly generated by PKC-dependent activation of NADPH oxidase","volume":"118","author":"Asano","year":"2012","journal-title":"J. Pharmacol. Sci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1680","DOI":"10.1093\/chromsci\/bmv071","article-title":"Determination of rutin and isoquercetin contents in Hibisci mutabilis folium in different collection periods by HPLC","volume":"53","author":"Liu","year":"2015","journal-title":"J. Chromatogr. Sci."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"529","DOI":"10.1021\/tx3004906","article-title":"Metabolic activation of polycyclic aromatic hydrocarbons and aryl and heterocyclic amines by human cytochromes P450 2A13 and 2A6","volume":"26","author":"Shimada","year":"2013","journal-title":"Chem. Res. Toxicol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1016\/S0891-5849(02)01289-3","article-title":"The cytotoxic effect of volatile organic compounds of the gas phase of cigarette smoke on lung epithelial cells","volume":"34","author":"Pouli","year":"2003","journal-title":"Free Radic. Biol. Med."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Naik, P., Fofaria, N., Prasad, S., Sajja, R.K., Weksler, B., Couraud, P.O., Romero, I.A., and Cucullo, L. (2014). Oxidative and pro-inflammatory impact of regular and denicotinized cigarettes on blood brain barrier endothelial cells: Is smoking reduced or nicotine-free products really safe?. BMC Neurosci., 15.","DOI":"10.1186\/1471-2202-15-51"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"26576","DOI":"10.1074\/jbc.M112.372813","article-title":"Nicotine and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone binding and access channel in human cytochrome P450 2A6 and 2A13 enzymes","volume":"287","author":"DeVore","year":"2012","journal-title":"J. Biol. Chem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"698","DOI":"10.1124\/dmd.115.063305","article-title":"Suppression of pulmonary cyp2a13 expression by carcinogen-induced lung tumorigenesis in a Cyp2A13-humanized mouse model","volume":"43","author":"Liu","year":"2015","journal-title":"Drug Metab. Dispos. Biol. Fate Chem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"1341","DOI":"10.1124\/dmd.114.059188","article-title":"Generation and characterization of a novel Cyp2A13\u2014Transgenic mouse model","volume":"42","author":"Jia","year":"2014","journal-title":"Drug Metab. Dispos."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1093\/toxsci\/64.2.185","article-title":"Inhibition of human cytochrome P450 2E1 by nicotine, cotinine, and aqueous cigarette tar extract in vitro","volume":"64","author":"Bombick","year":"2001","journal-title":"Toxicol. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"6987","DOI":"10.1021\/jm060519r","article-title":"Synthetic inhibitors of cytochrome P-450 2A6: Inhibitory activity, difference spectra, mechanism of inhibition, and protein cocrystallization","volume":"49","author":"Yano","year":"2006","journal-title":"J. Med. Chem."},{"key":"ref_39","first-page":"131","article-title":"Toxicity mechanisms of cigarette smoke on mouse fetus mitochondria","volume":"14","author":"Naserzadeh","year":"2015","journal-title":"Iran. J. Pharm. Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1111\/cpr.12091","article-title":"Rapamycin inhibits acrolein-induced apoptosis by alleviating ROS-driven mitochondrial dysfunction in male germ cells","volume":"47","author":"He","year":"2014","journal-title":"Cell Prolif."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1016\/S0006-8993(01)02485-4","article-title":"Inhibition of caspase-3-like activity reduces glutamate induced cell death in adult rat retina","volume":"904","author":"Chen","year":"2001","journal-title":"Brain Res."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1007\/978-1-4615-1589-0_2","article-title":"The epidemiology of lung cancer","volume":"105","author":"Williams","year":"2001","journal-title":"Cancer Treat. Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1682","DOI":"10.1021\/tx100147z","article-title":"Potent mutagenicity of 3-methylindole requires pulmonary cytochrome P450-mediated bioactivation: A comparison to the prototype cigarette smoke mutagens B(a)P and NNK","volume":"23","author":"Weems","year":"2010","journal-title":"Chem. Res. Toxicol."}],"container-title":["International Journal of Environmental Research and Public Health"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1660-4601\/14\/10\/1221\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T18:47:12Z","timestamp":1760208432000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1660-4601\/14\/10\/1221"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,10,13]]},"references-count":43,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2017,10]]}},"alternative-id":["ijerph14101221"],"URL":"https:\/\/doi.org\/10.3390\/ijerph14101221","relation":{},"ISSN":["1660-4601"],"issn-type":[{"type":"electronic","value":"1660-4601"}],"subject":[],"published":{"date-parts":[[2017,10,13]]}}}