Molecular & Cellular Toxicology

The Korean Society of Toxicogenomics and Toxicoproteomics (대한독성유전 단백체학회)
권호 표지

DNA Single Strand Breaks of Perchloroethylene and Its Bio-degradation Products by Single Cell Gel Electrophoresis Assay in Mammalian Cell System

  • Published : 2005.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

Perchloroethylene (tetrachloroethylene, PCE), a dry cleaning and degreasing solvent, can enter ground-water through accidental leak or spills. PCE can be degraded to trichloroethylene (TCE), 1, 1-dichloroethylene (DCE) and vinyl chloride (VC) as potential bio-product. These compounds have been reported that they can cause clinical diseases and cytotoxicity. However, only a little genotoxic information of these compounds has been known. In this study, we investigated DNA single strand breaks of PCE, TCE, DCE and VC by single cell gel electrophoresis assay, (comet assay) which is a sensitive, reliable and rapid method for DNA single strand breaks with mouse lymphoma L5178Y cells. From these results, $37.5\;{\mu}g/ml$ of PCE, $189\;{\mu}g/ml$ of TCE and $56.4\;{\mu}g/ml$ of DCE were revealed significant DNA damages in the absence of S-9 metabolic activation system meaning direct-acting mutagen. And in the presence of S-9 metabolic activation system, $41.5\;{\mu}g/ml$ of PCE, $328.7\;{\mu}g/ml$ of TCE and $949\;{\mu}g/ml$ of DCE were induced significant DNA damage. In the case of VC, it was revealed a significant DNA damage in the presence of S-9 metabolic activation system. Therefore, we suggest that chloroethylene compounds (PCE, TCE, DCE and VC) may be induced the DNA damage in a mammalian cell.

Keywords

References

  1. NIOSH, Cincinnati, OH. NIOSH. (National Institute for Occupational Exposure and Health). National Occupational Exposure Survey. 1981-1983 (1994)
  2. IARC. (International Agency for Research on Cancer). Dry cleaning, some chlorinated solvents used in dry cleaning and other industrial chemicals. IARC Monographs on the Carcinogenic Risks to Humans. 63, 33-223 (1995)
  3. Kilburn, K.H. Is neurotoxicity associated with environmental trichloroethylene (TCE). Arch. Environ. Health. 57, 113-120 (2002) https://doi.org/10.1080/00039890209602925
  4. Kuo, H.W. et al. VOC concentration in Taiwan's household drinking water. Sci. Total Environ. 208, 41-47 (1997) https://doi.org/10.1016/S0048-9697(97)00274-X
  5. Nelson, M.J.K., Montgomery, S.O., O'Neill, E.J. & Pritchard, P.H. Aerobic metabolism of trichloroethylene by a bacterial isolate. Appl. Environ. Microbiol. 52, 383-384 (1986)
  6. Coleman, N.V., Mattes, T.E., Gossett, J.M. & Spain, J.C. Biodegradation of cis-dichloroethene as the sole carbon source by a b-proteobacterium. Appl. Environ. Microbiol. 68, 2726-2730 (2002a) https://doi.org/10.1128/AEM.68.6.2726-2730.2002
  7. Coleman, N.V., Mattes, T.E., Gossett, J.M. & Spain, J.C. Phylogenetic and kinetic diversity of aerobic vinyl chloride-assimilating bacteria from contaminated sites. Appl. Environ. Microbiol. 68, 6162-6171 (2002b) https://doi.org/10.1128/AEM.68.12.6162-6171.2002
  8. Ryoo, D., Shim, H., Canada, K., Barbieri, P. & Wood, T.K. Aerobic degradation of tetrachloroethylene by toluene-o-xylene monooxygenase of Pseudomonas stutzeri OX1. Nature. Biotechnol. 18, 775-778 (2000) https://doi.org/10.1038/77344
  9. van Hylckama Vlieg, J.E.T. & Janssen, D.B. Formation and detoxification of reactive intermediates in the metabolism of chlorinated ethenes. J. Biotechnol. 85, 81-102 (2001) https://doi.org/10.1016/S0168-1656(00)00364-3
  10. Ferroni, C. et al. I. Neurobehavioral and neuroendocrine effects of occupational exposure to perchloroethylene. Neurotoxicology. 13, 243-248 (1992)
  11. Briving, C., et al. Chronic effects of perchloroethylene and trichloroethylene on the gerbil brain amino acids and glutathione. Neurotoxicology. 7(1), 101- 108 (1986)
  12. Lash, L.H. & Parker, J.C. Hepatic and renal toxicities associated with perchloroethylene. Pharmacol. Rev. 53(2), 177-208 (2001)
  13. Lash, L.H. et al. Renal and hepatic toxicity of trichloroethylene and its glutathione-derived metabolites in rats and mice: sex-, species-, and tissue-dependent differences. J. Pharmacol. Exp. Therapeut. 297(1), 155-164 (2001)
  14. Coleman, W.E., Lingg, R.D., Melton, R.G. & Kopfler, F.C. The occurrence of volatile organics in five drinking water supplies using gas chromatography/mass spectrophotometry, in Identification and Analysis of Organic Pollutants in Water (Keith LH ed) Ann. Arbor. Science Publishers, Ann. Arbor. MI. 305-327 (1976)
  15. Forkert, P.G., Stringer, V. & Troughton, K.M. Pulmonary toxicity of 1,1-dichloroethylene: correlation of early changes with covalent binding. Can. J. Physiol. Pharmacol. 64, 112-121 (1986) https://doi.org/10.1139/y86-017
  16. IARC. Overall evaluations of carcinogenicity: an updating of IARC Monographs Volumes 1 to 42. IARC Monogr. Eval. Carcinog. Risk Hum. S7, 373-376 (1987)
  17. Anderson, D. & Plewa, M.J. The international comet assay workshop, Mutagenesis. 13, 67-73 (1998) https://doi.org/10.1093/mutage/13.1.67
  18. Fairbairn, D.W., Walburger, D.K., Fairbairn, J.J. & O'Neill, K.L. Key morphologic changes and DNA strand breaks in human lymphoid cells: discriminating apoptosis from necrosis. Scanning. 18, 407-416 (1996) https://doi.org/10.1002/sca.1996.4950180603
  19. Singh, N.P., McCoy, M.T., Tice, R.R. & Schneider, E.L. A simple technique for quantitation of low levels of DNA damage in individual cells, Exp. Cell Res. 175, 184-191 (1988) https://doi.org/10.1016/0014-4827(88)90265-0
  20. Speit, G. & Hartmann, A. The comet assay (singlecell gel test). A sensitive genotoxicity test for the detection of DNA damage and repair. Methods Mol. Biol. 113, 203-212 (1999)
  21. Tice, R.R., Andrews, P.W., Hirai, O. & Singh, N.P. The single cell gel (SCG) assay: an electrophoretic technique for the detection of DNA damage in individual cells. Adv. Exp. Med. Biol. 283, 157-164 (1991)
  22. Tice, R.R. et al. Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ. Mol. Mutagen. 35, 206-221 (2000) https://doi.org/10.1002/(SICI)1098-2280(2000)35:3<206::AID-EM8>3.0.CO;2-J
  23. Devaux, A., Personen, M. & Monod, G. Alkaline comet assay in rainbow trout hepatocytes. Toxicol. In Vitro. 11, 71-79 (1997) https://doi.org/10.1016/S0887-2333(97)00004-0
  24. Wilson, J.T., Pascoe, P.L., Parry, J.M. & Dixon, D.R. Evaluation of the comet assay as a method for the detection of DNA damage in the cells of a marine invertebrate, Mytilus edulis L. (Mollusca: Pelecypoda). Mutat. Res. 399, 87-95 (1998)
  25. Kamer, I. & Rinkevich, B. In vitro application of the comet assay for aquatic genotoxicity: considering a primary culture versus a cell line. Toxicol. In Vitro. 16, 177-184 (2002) https://doi.org/10.1016/S0887-2333(01)00118-7
  26. Pradrangi, R., Petras, M., Ralph, S. & Vrzoc, M. Alkaline single cell gel (comet) assay and genotoxicity monitoring using bullheads and carp. Environ. Mol. Mutag. 26, 345-356 (1995) https://doi.org/10.1002/em.2850260411
  27. Mitchelmore, C.L., Birmelin, C., Chipman, J.K. & Livingstone, D.R. Detection of DNA strand breaks in mussel (Mytilus edulis L.) digestive gland cells using the 'comet' assay. Ecotoxicol. Environ. Saf. 41, 51- 58 (1998) https://doi.org/10.1006/eesa.1998.1666
  28. Ryu, J.C., Kim, H.T. & Kim, Y.J. Studies on DNA single strand break of seven phthalate analogues in mouse lymphoma cells. Environ. Mutagens & Carcinogens. 22(3), 164-168 (2002)
  29. Ryu, J.C. et al. Recent advanced toxicological methods for environmental hazardous chemicals. Kor. J. Environ. Toxicol. 14, 1-12 (1999)
  30. Cerna, M. & Kypenova, H. Mutagenic activity of chloroethylenes analysed by screening system tests. Mutat. Res. 46, 214-215 (1977)
  31. National cancer institute/national toxicology program, U.S. department of health and human services, NCI/NTP carcinogenesis technical report series, TR- 311 Y86
  32. National cancer institute/national toxicology program, U.S. department of health and human services, NCI/NTP carcinogenesis technical report series, TR- 273 Y88
  33. Zeiger, E., Anderson, B., Haworth, S., Lawlor, T. & Mortelmans, K. Salmonella mutagenicity tests: V. Results from the testing of 311 chemicals. Environ. Mol. Mutagen. 19(21), 2-141 (1992) https://doi.org/10.1002/em.2850190603
  34. McCann, J., Choi, E., Yamasaki, E. & Ames, B.N. Detection of carcinogens as mutagens in the Salmonella microsome test: assay of 300 chemicals. Proc. Natl. Acad. Sci. 72(12), 5135-5139 (1975) https://doi.org/10.1073/pnas.72.12.5135
  35. Zhu, S.M. et al. DNA damages of liver cells and expressions of DNA damage repair genes in rats exposed to vinyl chloride monomer. Zhonghua Lao Dong Wei Sheng Zhi Ye Bing Za Zhi. 22(3), 173-175 (2004)