Browse > Article

Genotoxic Effects of Diesel Exhaust Particle Extract in NIH/3T3 Cells  

Heo Chan (강원대학교 약학대학)
Kim Nam Yee (강원대학교 약학대학)
Chung Kyu-Hyuek (성균관대학교 약학대학)
Moon Chang-Kiu (서울대학교 약학대학)
Heo Moon Young (강원대학교 약학대학)
Publication Information
Environmental Analysis Health and Toxicology / v.19, no.4, 2004 , pp. 335-344 More about this Journal
Abstract
Diesel exhaust particle (<2.5 ${\mu}{\textrm}{m}$, DEP$_{2.5}$) is known to be probarbly carcinogenic (IARC group 2A). DEP$_{2.5}$ contains organic compounds such as polycyclicaromatic hydrocarbon (PAH), heterocyclic compounds, phenols, and nitroarenes. Reactive oxygen species (ROS) are generated by DEP$_{2.5}$ without any biological activation system. Therefore, an alternative mechanism by which DEP$_{2.5}$ could be carcinogenic is known by the generation of oxidative DNA damage. The aim of this study was to investigate genotoxic effects of DEP$_{2.5}$ using single cell gel electrophoresis. In order to evaluate the mechanisms of DEP$_{2.5}$ genotoxicity, the rat micro-some mediated and DNA repair enzyme treated comet assays together with routine comet assay were performed. DEP$_{2.5}$ was collected from diesel engine bus and dichloromethane extract was obtained. The organic extract of DEP$_{2.5}$ revealed DNA damage itself in NIH/3T3 cells. And it showed both oxidative and microsome mediated DNA damages. Vitamin C as an model antioxidant reduced DNA damage in endonuclase III treated comet assay. One of flavonoid, galangin as a CYP1A1 inhibitor reduced DNA damage in the presence of S-9 mixture. Our results show that DEP$_{2.5}$ are genotoxic and a great source of oxidative stress, but antioxidants can significantly reduce oxidative DNA damages. And DEP$_{2.5}$ may contain indirect mutagens which can be inhibited by CYP inhibitors.d by CYP inhibitors.
Keywords
diesel exhaust particle; single cell gel electrophoresis; reactive oxygen species (ROS); antioxidant; CYP1A1 inhibitor;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Cole SP. Rapid chemosensitivity testing of human lung tumor cells using the MTT assay, Cancer Chemother. Pharmacol. 1986; 17: 259-263
2 Iwai K, Adachi S, Takahashi M, Moller L, Udagawa T, Mizuno S and Sugawara I. Oxidative DNA damages and lung cancer, Env. Res. 2000; 84(3): 255-264   DOI   ScienceOn
3 Moller P, Daneshvar B, Loft S, Wallin H, Poulsen HE, Autrup H, Ravn-Haren G and Dragsted LO. Oxidative DNA damage in vitamin C-supplemented guinea pigs after intratracheal instillation of diesel exhaust particles, Toxicol. Appl. Pharmacol. 2003; 189(1): 39-44   DOI   ScienceOn
4 Pelkonen 0 and Nebert DW. Metabolism of polycyclic aromatic hydrocarbons: etiologic role in carcinogenesis, Pharmacol. Rev. 1982; 34(2): 189-222
5 Scheepers PT, Coggon D, Knudsen LE, Anzion R, Autrup H, Bogovsld S, Bos RP, Dahmann D, Farmer P, Martin EA, Micka V, Muzyka V, Neumann HG, Poole J, Schmidt Ott A, Seiler F, Volf J and Zwirner-Baier I. BIOMarkers for occupational diesel exhaust exposure monitoring (BIOMODEM)--a study in underground mining, Toxicol. Lett. 2002, 134(1-3): 305-317   DOI   ScienceOn
6 Schins RP. Mechanisms of genotoxicity of particles and fibers, Inhal. Toxicol. 2002; 14(1): 57-78   DOI   PUBMED   ScienceOn
7 Gelboin HV. Benzo[a]Pyrene metabolism, activation and carcinogenesis: role and regulation of mixed-function oxidases and related enzymes, Physiol. Rev. 1980; 60(4): 1107-1166   DOI
8 Mauderly JL. Lung overload: the dilemma and opportunities for resolution, Inhalation Toxicology 1996; 8: 1-28   DOI
9 Quadri SA, Qadri AN, Hahn ME, Mann KK and Sherr DH. The bioflavonoid galangin blocks aryl hydrocarbon receptor activation and polycyclic aromatic hydrocarbon-induced pre-B cell apoptosis, Mol. Pharmacol. 2000; 58(3): 515-525   DOI
10 International Agency for Research on Cancer(IARC). Diesel and gasoline engine exhausts and some nitroarenes. In: Monographs on the Evaluation of Carcinogenic Risks to Humans, World Health Organization. IARC, Lyon, France. 1989; 46
11 McClellan RO. Lung cancer in rats from prolonged exposure to high concentrations of carbonaceous particles: implication for human risk assessment, Inhalation Toxicology 1996; 8: 193-226
12 Lewtas J and Gallagher J. Complex mixtures of urban air pollutants: identification and comparative assessment of mutagenic and tumorigenic chemicals and emission sources, IARC Sci. Publ. 1990; 104: 252-260
13 Don Porto Carero A, Hoet PH, Verschaeve L, Schoeters G and Nemery B. Genotoxic effects of carbon black Particles, diesel exhaust particles, and urban air paiticulates and their extracts on a human alveolar epithelial cell line (A549) and a human monocytic cell line (THP-l), Environ. Mol. Mutagen 2001; 37(2): 155-163   DOI   ScienceOn
14 Singh NP, McCoy MT, Tice RR and Schneider EL. A simple technique for quantitation of low levels of DNA damage in individual cells, Experimental Cell Research 1988; 175:184-191   DOI   ScienceOn
15 Pohjola SK, Lappi M, Honkanen M, Rantanen L and Savela K. DNA binding of polycyclic aromatic hydrocarbons in a human bronchial epithelial cell line treated with diesel and gasoline particulate extracts and benzo[a]Pyrene, Mutagenesis 2003; 18(5): 429-438   DOI   ScienceOn
16 환경부. 환경백서, 2000
17 Farombi EO, Moller P and Dragsted LO. Ex-vivo and in vitro protective effects of kolaviron against Oxygen-derived radical-induced DNA damage and oxidative stress in human lymphocytes and rat liver cells, Cell Biol. Toxicol. 2004; 20(2): 71-82   DOI   ScienceOn
18 Health Effects Institute (HEI). Program summary: Research on Diesel Exhaust, Health Effects Institute. Cambridge, MA. 1999
19 National Institute for Occupational Safety and Health (NIOSH). Carcinogenic Effects of Exposure to Diesel Exhaust. NIOSH Current Intelligence Bulletin 50. DHHS (NIOSH), Centers for Disease Control. Atlanta, GA. 1988
20 Muller AK, Farombi EO, Moller P, Autrup HN, Vogel U, Wallin H, Dragsted LO, Loft S and Binderup ML. DNA damage in lung after oral exposure to diesel exhaust particles in Big Blue rats, Mutat. Res. 2004; 550(1-2): 123-132   DOI
21 Greenwell LL, Moreno T, Jones TP and Richards RJ. Particle-induced oxidative damage is ameliorated by pulmonary antioxidants, Free Radic. Biol. Med. 2002; 32(9): 898-905   DOI   ScienceOn
22 Annas A, Brittebo E and Hellman B. Evaluation of benzo (a)Pyrene-induced DNA damage in human endothelial cells using alkaline single cell gel electrophoresis, Mutat Res. 2000; 471(1-2): 145-155   DOI
23 Dybdahl M, Risom L, Moller P, Autrup H, Wallin H, Vogel U, Bornholdt J, Daneshvar B, Dragsted LO, Weimann A, Poulsen HE and Loft S. DNA adduct formation and oxidative stress in colon and liver of Big Blue rats after dietary exposure to diesel particles, Carcinogenesis 2003; 24(11): 1759-1766   DOI   ScienceOn
24 International Agency for Research on Cancer. IARC monographs on the evaluation of carcinogenic risks to humans. Diesel and gasoline engine exhausts and some nitroarenes. International Agency for Research on Cancer. IARC Monogr. Eval. Carcinog. Risks Hum. 1989; 46: 1-458
25 Baulig A, Garlatti M, Bonvallot V, Marchand A, Barouki R, Marano F and Baeza-Squiban A. Involvement of reactive oxygen species in the metabolic pathways triggered by diesel exhaust particles in human airway epithelial cells, Am J Physiol Lung CelI Mol. Physiol. 2003; 285(3): 671-679   DOI
26 Heo MY, Sohn SJ and Au WW. Anti-genotoxicity of galangin as a cancer chemopreventive agent candidate, Mutation Research 2001; 488: 135-150   DOI   ScienceOn
27 Le Marchand L, Murphy SP, Hankin JH, Wilkens LR and Kolonel LN. Intake of flavonoids and lung cancer, J. Natl. Cancer lnst. 2000; 92(2): 154-160   DOI   ScienceOn
28 U.S. Environmental Protection Agency (USEPA) Health Assessment Document for Diesel Emissions: SAB Review Draft. EPA/600/8-90/057D. National Center for Environmental Assessment, Washington DC. 1999
29 Das M, Khan WA, Asokan P, Bickers DR and Mukhtar H. Inhibitio of polycyclic aromatic hydrocarbon-DNA adduct formation in epidermis and lungs of SENCAR mice by naturally occurring plant phenols, Cancer Res. 1987; 47(3): 767-773
30 Duez P, Dehon G and Dubois J. Validation of raw data measurements in the comet assay, Talanta 2004; 63(4): 879-886   DOI   ScienceOn
31 World Health Organization (WHO) Diesel fuel and exhaust emissions. International Program on Chemical Safety, Environmental Health Criteria 171, Geneva, Switzerland. 1996
32 Kato A, Nagai A and Kagawa J. Morphological changes in rat lung after long-term exposure to diesel emissions, Inhalation Toxicology 2000; 12: 469-490   DOI   ScienceOn
33 Health Effects Institute (HEI). Diesel exhaust: a critical analysis of emissions, exposure and health effects. A Special Report of the Institute's Diesel Working GrouP, Health Effects Institute. Cambridge, MA. 1995
34 Li N, Wang M, Oberley TD, Sempf JM and Nel AE. Comparison of the pro-oxidative and proinflammatory effects of organic diesel exhaust particle chemicals in bronchial epithelial cells and macrophages, J. Immunol. 2002; 169(8): 4531-4541   DOI
35 Edenharder R, Krieg H, Kottgen V and Platt KL. Inhibition of clastogenicity of benzo[a]Pyrene and ofits trans-7,8-dihydrodiol in mice in vivo by fruits, vegetables, and flavonoids, Mutat. Res. 2003; 537(2): 169-181   DOI
36 Heo MY, Yu KS, Kim KH, Kim HP and Au WW. Anticlastogenic effect of flavonoid against mutagen-induced micronuclei in mice, Mutation Research 1992; 284: 243-249
37 Kagawa J. Health effects of diesel exhaust emissions--amixture of air pollutants of worldwide concern, Toxicology 2002; 27: 349-353