Nicotine Component of Cigarette Smoke Extract (CSE) Decreases the Cytotoxicity of CSE in BEAS-2B Cells Stably Expressing Human Cytochrome P450 2A13
| Autor: | Rong Xia, Zhan Zhang, Minghui Ji, Na Li, Shoulin Wang, Chao Wang, Yudong Zhang |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Nitrosamines Cell Survival Health Toxicology and Mutagenesis Poly ADP ribose polymerase lcsh:Medicine Apoptosis Bronchi Cell Count Article Cell Line Flow cytometry Nicotine 03 medical and health sciences Tandem Mass Spectrometry BEAS-2B cells parasitic diseases medicine Humans Viability assay Cytotoxicity B cell bcl-2-Associated X Protein cigarette smoke extract medicine.diagnostic_test Caspase 3 Chemistry lcsh:R Public Health Environmental and Occupational Health Molecular biology nicotine cytochrome P450 2A13 cytotoxicity 030104 developmental biology CYP2A13 medicine.anatomical_structure Biochemistry Tobacco Smoke Pollution Aryl Hydrocarbon Hydroxylases medicine.drug |
| Zdroj: | International Journal of Environmental Research and Public Health; Volume 14; Issue 10; Pages: 1221 International Journal of Environmental Research and Public Health, Vol 14, Iss 10, p 1221 (2017) International Journal of Environmental Research and Public Health |
| ISSN: | 1660-4601 |
| DOI: | 10.3390/ijerph14101221 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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