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Measurement of DNA Damage with Fpg/Endo III FLARE Assay and Real Time RT-PCR in SD Rats Exposed to Cumene  

Kim, Soo-Jin (Laboratory of Occupational Toxicology, Chemical Safety & Health Research Center, Occupational Safety & Health Research Institute, KOSHA)
Rim, Kyung-Taek (Laboratory of Occupational Toxicology, Chemical Safety & Health Research Center, Occupational Safety & Health Research Institute, KOSHA)
Lee, Seong-Bae (Laboratory of Occupational Toxicology, Chemical Safety & Health Research Center, Occupational Safety & Health Research Institute, KOSHA)
Kim, Hyeon-Yeong (Laboratory of Occupational Toxicology, Chemical Safety & Health Research Center, Occupational Safety & Health Research Institute, KOSHA)
Publication Information
Molecular & Cellular Toxicology / v.4, no.3, 2008 , pp. 211-217 More about this Journal
Abstract
To clarify the DNA damage from reactive oxygen species, we measured the DNA damage through Fpg/Endo III FLARE (Fragment Length Analysis with Repair Enzyme) assay and real time RT-PCR. The 80 SD rats assigned to 4 dose groups exposed to cumene vapor for 90 days. With Fpg/Endo III FLARE assay in hepatocytes, we found the OTM (Olive Tail Moment) and TL (Tail Length) significantly increased in no-enzyme treated and Fpg-treated control and 8 ppm groups with 28 days exposure. In Endo III-treated 8 ppm group, significantly increased the values with 90 days exposure. With lymphocytes, it was founded the values significantly increased in no-enzyme treated 800 ppm group in 28 and 90 days. It was significantly increased in Endo III-treated 80 ppm for 28 days and 800 ppm for 90 days. From the above findings, FLARE assay was suggested as being available as a biological marker for DNA damage induced by cumene exposure in SD rats. And we used real time RT-PCR for the OGG1 mRNA expression, it had dose-dependent biologic effects in 1 day exposure, but decrease the levels of rOGG1 mRNA. Our findings provide evidence that cumene exposure may cause suppression of rOGG1 in the rat hepatocytes or lymphocytes.
Keywords
Cumene; Fpg/Endo III FLARE assay; Real time RT-PCR; OGG1;
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  • Reference
1 World Health Organization (WHO)/International Programme on Chemical Safety (IPCS). Concise international chemical assessment document No. 18. Cumene p. 4, Geneva, Switzerland (1999)
2 Jackson, J. et al. Test Rule Support Document Cumene Syracuse Res Corp p 170 (1985)
3 Chemical Manufacturers Association (CMA). Cumene Program Panel: Industrial Hygiene survey. (1985)
4 American Petroleum Institute (API). Letter to TSCA Interagency Testing Committee USEPA (1984)
5 Cocheo, V. et al. Rubber manufacture: Sampling and identification of volatile pollutants. Amer Ind Hyg Assoc J 44:521-527 (1983)   DOI
6 National Institute of Occupational Safety & Health (NIOSH). National Occupational Exposure Survey (NOES) (1983)
7 Schulz, R. C. et al. Kirk-Othmer Encycl Chem Technol. 4th ed. NY, NY: John Wiley and Sons 7:735(1993)
8 Droge, W. Free radicals in the physiological control of cell function. Physiol Rev 82:47-95 (2002)   DOI
9 Hazra, T. K., Izumi, T., Maidt, L., Floyd, R. A. & Mitra, S. The presence of two distinct 8-oxoguanine repair enzymes in human cells: their potential complementary roles in preventing mutation. Nucleic Acids Res 26:5116-5122 (1999)   DOI   ScienceOn
10 Collins, A. R., Duthie, S. J. & Dobson, V. L. Direct enzymatic detection of endogenous oxidative base damage in human lymphocyte DNA. Carcinogenesis 14:1733-1735 (1993)   DOI   ScienceOn
11 Collins, A. R., Dusinska, M., Gedik, C. M. & Stetina, R. Oxidative damage to DNA: do we have a reliable biomarker? Environ Health Perspect 104:465-469 (1996)   DOI
12 Duthie, S. J., Ma, A., Ross, M. A. & Collins, A. R. Antioxidant supplementation decreases oxidative DNA damage in human lymphocytes. Cancer Res 56: 1291-1295 (1996)
13 Yarosh, D. B., Rice, M., Day, R. S. 3rd, Foote, R. S. & Mitra, S. O6-Methylguanine-DNA methyltransferase in human cells. Mutation Research 131:27-36 (1984)   DOI   ScienceOn
14 Samson, L. & Cairns, J. A new pathway for DNA repair in Escherichia coli. Nature 267:281-283 (1977)   DOI   ScienceOn
15 Ikushima, T. Radioadaptive response: Characterization of a cytogenetic repair induced by low-level ionizing radiation in cultured Chinese hamster cells. Mutation Research Letters 227:241-246 (1989)   DOI   ScienceOn
16 Doull, J., Klassen, C. D. & Amdur, M. D. Casarett and Doull's Toxicology. 3rd ed. New York: Macmillan Co. Inc. p. 644 (1986)
17 U.S. Environmental Protection Agency's Integrated Risk Information System (IRIS) for Cumene (98-82- 8). Available from: http://www.epa.gov/ngispgm3/iris on the Substance File list (2000)
18 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)   DOI   ScienceOn
19 Scott, B. R. A biological-based model that links genomic instability, bystander effects, and adaptive response. Mutat Res 568:129-143 (2004)   DOI
20 Kasai, H. et al. Formation of 8-hydroxyguanine moiety in cellular DNA by agents producing oxygen radicals and evidence for its repair. Carcinogenesis 7:1849-1851 (1986)   DOI   ScienceOn
21 Ostling, O. & Johanson, K. J. Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells. Biochem Biophys Res Commun 123:291-298 (1984)   DOI   ScienceOn
22 Shibutani, S., Takeshita, M. & Grollan, A. P. Insertion of specific bases during DNA synthesis past the oxidation-damaged base 8-oxo dG. Nature 349:431-434 (1991)   DOI   ScienceOn
23 Phoa, N. & Epe, B. Influence of nitric oxide on the generation and repair of oxidative DNA damage in mammalian cells. Carcinogenesis 23:469-475 (2002)   DOI   ScienceOn
24 Olive, P. L. & Banath, J. P. Detection of DNA double -strand breaks through the cell cycle after exposure to X-rays, bleomycin, etoposide and 125IdUrd. Int J Radiat Biol 64:349-358 (1993)   DOI
25 Thelestam, M., Curvall, M. & Enzell, C. R. Effect of tobacco smoke compounds on the plasma membrane of cultured human lung fibroblates. Toxicology 15:203-217 (1980)   DOI   ScienceOn
26 Kasai, H. & Nishimura, S. Hydroxylation of deoxyguanosine at the C-8 position by ascorbic acid and other reducing agents. Nucleic Acids Res 12:2137-2145 (1984)   DOI   ScienceOn
27 Covallo, O. et al. Evaluation of oxidative damage and inhibition of DNA repair in an in vitro study of nickel exposure. Toxicology In Vitro 17:603-607 (2003)   DOI   ScienceOn
28 Dhenaut, A., Boiteux, S. & Radicella, J. P. Characterization of the hOGG1 promoter and its expression during the cell cycle. Mutat Res 461:109-118 (2000)   DOI   ScienceOn
29 Mackison, F. W., Stricoff, R. S. & Partridge, L. J. Jr. Occupational Health Guidelines for Chemical Hazards. DHHS(NIOSH) 3:81-123 (1981)
30 Shah, J. J. & Singh, H. B. Distribution of volatile organic chemicals in outdoor and indoor air. Environ Sci Technol 22:1381-1388 (1988)   DOI   ScienceOn
31 Fairbairn, D. W., Olive, P. L. & O'Neill, K. L. The comet assay: A comprehensive review. Mutat Res 339: 37-59 (1995)   DOI   ScienceOn
32 Kruszewski, M., Wojewodzka, M., Iwanenko, T., Collins, A. R. & Szumiel, I. Application of the comet assay for monitoring DNA damage in workers exposed to chronic low-dose irradiation. II. Base damage. Mutat Res 416:37-57 (1998)   DOI
33 Potts, R. J. Watkin, R. D. & Hart, B. A. Cadmium exposure down-regulates 8-oxoguanine DNA glycosylase expression in rat lung and alveolar epithelial cells. Toxicology 184:189-202 (2003)   DOI   ScienceOn
34 Pant, M. C. et al. Mechanisms of suppression of neoplastic transformation in vitro by low doses of low LET radiation. Carcinogenesis 24:1961-1965 (2003)   DOI   ScienceOn
35 Kamiya, H. et al. c-H-ras containing 8-hydroxyguanine at codon 12 induces point mutations at the modified and adjacent position. Cancer Res 52:3483-3485 (1992)
36 Yu, I. J. et al. Inflammatory and genotoxic responses during 30-day welding-fume exposure period. Toxicol Lett 154:105-115 (2004)   DOI   ScienceOn
37 Abalea et al. Iron-induced oxidative DNA damage and its repair in primary rat hepatocyte culture. Carcinogenesis 19:1053-1059 (1998)   DOI   ScienceOn
38 Asmuss, M., Mullenders, L. H., Eker, A. & Hartwig, A. Differential effects of toxic metal compounds on the activities of Fpg and XPA, two zinc finger proteins involved in DNA repair. Carcinogenesis 21:2097-2104 (2000)   DOI   ScienceOn
39 Kasai, H. Analysis of a form of oxidative DNA damage, 8-hydroxy-2′-deoxyguanosine, as a marker of cellular oxidative stress during carcinogenesis. Mutat Res 387:147-163 (1997)
40 Marnett, L. J. Oxiradicals and DNA damage. Carcinogenesis 21:361-370 (2000)   DOI   ScienceOn
41 Asami, S. et al. Effects of forced and spontaneous exercise on 8-hydroxydeoxyguanosine levels in rat organs. Biochem Biophys Res Commun 243:678-682 (1998)   DOI   ScienceOn
42 Lewis, R. J. Sax's Dangerous Properties of Industrial Materials. (9th ed.) New York: Van Nostrand reinhold 1-3:938 (1996)
43 Lewis, R. J., Sr. Hawley's Condensed Chemical Dictionary. (12th ed.) New York: Van Nostrand rheinhold Co. p. 329 (1993)
44 Collins, A. R., Dobson, V., Dusinska, M., Kennedy, G. & Stetina, R. The comet assay: what can it really tell us? Mutat Res 375:183-193 (1997)   DOI   ScienceOn
45 Hartwig, A. Carcinogenicity of metal compounds: possible role of DNA repair inhibition. Toxicol Lett 102-103:235-239 (1998)   DOI   ScienceOn
46 Tice, R. R. et al. Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen 35:206-21 (2000)   DOI   ScienceOn
47 Maeng, S. H. et al. Changes of 8-OH-dG levels in DNA and its base excision repair activity in rat lungs after inhalation exposure to hexavalent chromium. Mutat Res 539:109-116 (2003)   DOI
48 Jekinson, A. M., Collins, A. R., Duthie, S. J., Wahle, K. W. J. & Duthie, G. G. The effect of increased intakes of polyunsaturated fatty acids and vitamin E on DNA damage in human lymphocytes. FASEB J 13: 2138-2142 (1999)   DOI
49 Documentation of the Threshold Limit Values and Biological Exposure Indices, Cincinnati, OH (5th ed.). American Conference of Governmental Industrial Hygienists (ACGIH). p. 151 (1986)