Comet Assay Evaluation of Six Chemicals of Known Genotoxic Potential in Rats

Overview

Journal Mutat Res Genet Toxicol Environ Mutagen

Publisher Elsevier

Specialties Environmental Health
Genetics
Toxicology

Date 2015 Jul 28

PMID 26212309

Citations 8

Authors

Cheryl A Hobbs

Leslie Recio

Michael Streicker

Molly H Boyle

Jin Tanaka

Atsushi Shiga

Kristine L Witt

Affiliations

Soon will be listed here.

Abstract

As a part of an international validation of the in vivo rat alkaline comet assay (comet assay) initiated by the Japanese Center for the Validation of Alternative Methods (JaCVAM) we examined six chemicals for potential to induce DNA damage: 2-acetylaminofluorene (2-AAF), N-nitrosodimethylamine (DMN), o-anisidine, 1,2-dimethylhydrazine dihydrochloride (1,2-DMH), sodium chloride, and sodium arsenite. DNA damage was evaluated in the liver and stomach of 7- to 9-week-old male Sprague Dawley rats. Of the five genotoxic carcinogens tested in our laboratory, DMN and 1,2-DMH were positive in the liver and negative in the stomach, 2-AAF and o-anisidine produced an equivocal result in liver and negative results in stomach, and sodium arsenite was negative in both liver and stomach. 1,2-DMH and DMN induced dose-related increases in hedgehogs in the same tissue (liver) that exhibited increased DNA migration. However, no cytotoxicity was indicated by the neutral diffusion assay (assessment of highly fragmented DNA) or histopathology in response to treatment with any of the tested chemicals. Therefore, the increased DNA damage resulting from exposure to DMN and 1,2-DMH was considered to represent a genotoxic response. Sodium chloride, a non-genotoxic non-carcinogen, was negative in both tissues as would be predicted. Although only two (1,2-DMH and DMN) out of five genotoxic carcinogens produced clearly positive results in the comet assay, the results obtained for o-anisidine and sodium arsenite in liver and stomach cells are consistent with the known mode of genotoxicity and tissue specificity exhibited by these carcinogens. In contrast, given the known genotoxic mode-of-action and target organ carcinogenicity of 2-AAF, it is unclear why this chemical failed to convincingly increase DNA migration in the liver. Thus, the results of the comet assay validation studies conducted in our laboratory were considered appropriate for five out of the six test chemicals.

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References

Ashby J . The genotoxicity of sodium saccharin and sodium chloride in relation to their cancer-promoting properties. Food Chem Toxicol. 1985; 23(4-5):507-19. DOI: 10.1016/0278-6915(85)90145-0. View

Shephard S, Sengstag C, Lutz W, Schlatter C . Mutations in liver DNA of lacI transgenic mice (Big Blue) following subchronic exposure to 2-acetylaminofluorene. Mutat Res. 1993; 302(2):91-6. DOI: 10.1016/0165-7992(93)90009-k. View

Tinwell H, Stephens S, Ashby J . Arsenite as the probable active species in the human carcinogenicity of arsenic: mouse micronucleus assays on Na and K arsenite, orpiment, and Fowler's solution. Environ Health Perspect. 1991; 95:205-10. PMC: 1568403. DOI: 10.1289/ehp.9195205. View

Sekihashi K, Yamamoto A, Matsumura Y, Ueno S, Watanabe-Akanuma M, Kassie F . Comparative investigation of multiple organs of mice and rats in the comet assay. Mutat Res. 2002; 517(1-2):53-75. DOI: 10.1016/s1383-5718(02)00034-7. View

. Re-evaluation of some organic chemicals, hydrazine and hydrogen peroxide. IARC Monogr Eval Carcinog Risks Hum. 1999; 71 Pt 1, Pt 2, Pt 3:1-1554. PMC: 7681305. View

Rothfuss A, ODonovan M, De Boeck M, Brault D, Czich A, Custer L . Collaborative study on fifteen compounds in the rat-liver Comet assay integrated into 2- and 4-week repeat-dose studies. Mutat Res. 2010; 702(1):40-69. DOI: 10.1016/j.mrgentox.2010.07.006. View

Xie Y, Liu J, Benbrahim-Tallaa L, Ward J, Logsdon D, Diwan B . Aberrant DNA methylation and gene expression in livers of newborn mice transplacentally exposed to a hepatocarcinogenic dose of inorganic arsenic. Toxicology. 2007; 236(1-2):7-15. PMC: 2465467. DOI: 10.1016/j.tox.2007.03.021. View

Kadirvel R, Sundaram K, Mani S, Samuel S, Elango N, Panneerselvam C . Supplementation of ascorbic acid and alpha-tocopherol prevents arsenic-induced protein oxidation and DNA damage induced by arsenic in rats. Hum Exp Toxicol. 2008; 26(12):939-46. DOI: 10.1177/0960327107087909. View

Ohsawa K, Furihata C, Mori M, Ikui E . Ability of N-methyl-N'-nitro-N-nitrosoguanidine, 4-nitroquinoline 1-oxide, dimethylnitrosamine, and NaCl to induce unscheduled DNA synthesis, stimulate replicative DNA synthesis, and produce DNA single-strand breaks in pyloric mucosa of rat stomach. Mutat Res. 1993; 287(2):307-19. DOI: 10.1016/0027-5107(93)90023-9. View

10.

Tsuda S, Matsusaka N, Madarame H, Ueno S, Susa N, Ishida K . The comet assay in eight mouse organs: results with 24 azo compounds. Mutat Res. 2000; 465(1-2):11-26. DOI: 10.1016/s1383-5718(99)00199-0. View

11.

Madle S, Dean S, Andrae U, Brambilla G, Burlinson B, Doolittle D . Recommendations for the performance of UDS tests in vitro and in vivo. Mutat Res. 1994; 312(3):263-85. DOI: 10.1016/0165-1161(94)00013-1. View

12.

Lambert I, Singer T, Boucher S, Douglas G . Detailed review of transgenic rodent mutation assays. Mutat Res. 2005; 590(1-3):1-280. DOI: 10.1016/j.mrrev.2005.04.002. View

13.

Mavournin K, BLAKEY D, Cimino M, Salamone M, Heddle J . The in vivo micronucleus assay in mammalian bone marrow and peripheral blood. A report of the U.S. Environmental Protection Agency Gene-Tox Program. Mutat Res. 1990; 239(1):29-80. DOI: 10.1016/0165-1110(90)90030-f. View

14.

. Bioassay of o-anisidine hydrochloride for possible carcinogenicity. Natl Cancer Inst Carcinog Tech Rep Ser. 1978; 89:1-149. View

15.

Rothfuss A, Honma M, Czich A, Aardema M, Burlinson B, Galloway S . Improvement of in vivo genotoxicity assessment: combination of acute tests and integration into standard toxicity testing. Mutat Res. 2010; 723(2):108-20. DOI: 10.1016/j.mrgentox.2010.12.005. View

16.

Ashby J, Lefevre P, Tinwell H, Brunborg G, Schmezer P, Pool-Zobel B . The non-genotoxicity to rodents of the potent rodent bladder carcinogens o-anisidine and p-cresidine. Mutat Res. 1991; 250(1-2):115-33. DOI: 10.1016/0027-5107(91)90168-n. View

17.

Takahashi M, Nishikawa A, Furukawa F, Enami T, Hasegawa T, Hayashi Y . Dose-dependent promoting effects of sodium chloride (NaCl) on rat glandular stomach carcinogenesis initiated with N-methyl-N'-nitro-N-nitrosoguanidine. Carcinogenesis. 1994; 15(7):1429-32. DOI: 10.1093/carcin/15.7.1429. View

18.

Stiborova M, Miksanova M, Sulc M, Rydlova H, Schmeiser H, Frei E . Identification of a genotoxic mechanism for the carcinogenicity of the environmental pollutant and suspected human carcinogen o-anisidine. Int J Cancer. 2005; 116(5):667-78. DOI: 10.1002/ijc.21122. View

19.

Watanabe M, Honda S, Hayashi M, Matsuda T . Mutagenic effects of combinations of chemical carcinogens and environmental pollutants in mice as shown by the micronucleus test. Mutat Res. 1982; 97(1):43-8. DOI: 10.1016/0165-1161(82)90018-8. View

20.

Soffritti M, Belpoggi F, Esposti D, Lambertini L . Results of a long-term carcinogenicity bioassay on Sprague-Dawley rats exposed to sodium arsenite administered in drinking water. Ann N Y Acad Sci. 2006; 1076:578-91. DOI: 10.1196/annals.1371.075. View