Environmental Analysis Health and Toxicology

The Korean Society of Environmental Health and Toxicology (환경독성보건학회)
권호 표지

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

디젤분진이 체세포에서의 DNA 손상에 미치는 영향

  • Published : 2004.12.01
Download PDF
⟨ Previous Next ⟩

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

References

  1. 환경부. 환경백서, 2000
  2. 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 https://doi.org/10.1016/S1383-5718(00)00127-3
  3. 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 https://doi.org/10.1152/ajplung.00419.2002
  4. Cole SP. Rapid chemosensitivity testing of human lung tumor cells using the MTT assay, Cancer Chemother. Pharmacol. 1986; 17: 259-263
  5. 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
  6. 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 https://doi.org/10.1002/em.1023
  7. Duez P, Dehon G and Dubois J. Validation of raw data measurements in the comet assay, Talanta 2004; 63(4): 879-886 https://doi.org/10.1016/j.talanta.2003.12.035
  8. 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 https://doi.org/10.1093/carcin/bgg147
  9. 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 https://doi.org/10.1016/S1383-5718(03)00078-0
  10. 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 https://doi.org/10.1023/B:CBTO.0000027916.61347.bc
  11. 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 https://doi.org/10.1152/physrev.1980.60.4.1107
  12. 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 https://doi.org/10.1016/S0891-5849(02)00782-7
  13. 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
  14. Health Effects Institute (HEI). Program summary: Research on Diesel Exhaust, Health Effects Institute. Cambridge, MA. 1999
  15. 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
  16. Heo MY, Sohn SJ and Au WW. Anti-genotoxicity of galangin as a cancer chemopreventive agent candidate, Mutation Research 2001; 488: 135-150 https://doi.org/10.1016/S1383-5742(01)00054-0
  17. 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
  18. 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
  19. 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 https://doi.org/10.1006/enrs.2000.4072
  20. Kagawa J. Health effects of diesel exhaust emissions--amixture of air pollutants of worldwide concern, Toxicology 2002; 27: 349-353
  21. 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 https://doi.org/10.1080/089583700402879
  22. 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
  23. 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 https://doi.org/10.4049/jimmunol.169.8.4531
  24. 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 https://doi.org/10.1093/jnci/92.2.154
  25. Mauderly JL. Lung overload: the dilemma and opportunities for resolution, Inhalation Toxicology 1996; 8: 1-28 https://doi.org/10.3109/08958379609005424
  26. 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
  27. 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 https://doi.org/10.1016/S0041-008X(03)00098-X
  28. 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 https://doi.org/10.1016/j.mrfmmm.2004.02.010
  29. 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
  30. Pelkonen 0 and Nebert DW. Metabolism of polycyclic aromatic hydrocarbons: etiologic role in carcinogenesis, Pharmacol. Rev. 1982; 34(2): 189-222
  31. 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 https://doi.org/10.1093/mutage/geg021
  32. 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 https://doi.org/10.1124/mol.58.3.515
  33. 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 https://doi.org/10.1016/S0378-4274(02)00195-9
  34. Schins RP. Mechanisms of genotoxicity of particles and fibers, Inhal. Toxicol. 2002; 14(1): 57-78 https://doi.org/10.1080/089583701753338631
  35. 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 https://doi.org/10.1016/0014-4827(88)90265-0
  36. 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
  37. World Health Organization (WHO) Diesel fuel and exhaust emissions. International Program on Chemical Safety, Environmental Health Criteria 171, Geneva, Switzerland. 1996