Validation of Photo-comet Assay as a Model for the Prediction of Photocarcinogenicity

  • Kim, Ji-Young (Laboratory of Genetic Toxicology, Korea Institute of Toxicology, KRICT) ;
  • Koh, Woo-Suk (Laboratory of Genetic Toxicology, Korea Institute of Toxicology, KRICT) ;
  • Lee, Mi-Chael (Department of Biology, College of Natural Sciences, University of Incheon)
  • 발행 : 2006.12.30

초록

Recent reports on the photocarcinogenicity and photogerotoxicity of many compounds led to an increasing awareness for the need of a standard approach to test for photogenotoxicity. The comet assay has been recently validated as a sensitive and specific test system for the quantification of DNA damage. Thus, the objectives of this study are to investigate the utility of photo-comet assay for detecting photo-mutagens, and to evaluate its ability to predict rodent photo-carcinogenicity. Photo-comet assays were performed using L5178Y $Tk^{+/-}$ mouse lymphoma cells on five test substances (8-methoxypsoralen, chlorpromazine, lomefloxacin, anthracene and retinoic acid) that demonstrated positive results in photocarcinogenicity tests. For the best discrimination between the test substance-mediated DNA damage and the undesirable DNA damage caused by direct UV absorption, a UV dose-response of the cells in the absence of the test substances was firstly fnalized. Out of 5 test substances, positive comet results were obtained for chlorpromazine, lomefloxacin, anthracene and retinoic acid while 8-methoxypsoralen found negative. An investigation into the predictive value of this photo-comet assay for determining the photocarcinogenicity showed that photo-comet assay has relatively high sensitivity. Therefore, the photo-comet assay with mammalian cells seems to be a good and sensitive predictor of the photocarcinogenic potential of new substances.

키워드

참고문헌

  1. Ball, P., Mandell, L. and Toillotson, G. (1999): Comparative tolerability of the newer fluoroquinolone antibacterials. Drug Safety, 21, 407-421 https://doi.org/10.2165/00002018-199921050-00005
  2. Blackburn, G.M. and Taussig, P.E. (1975): The photocarcinogenicity of anthracene: photochemical binding to deoxyribonucleic acid in tissue culture. Biochem. J., 149, 289-291 https://doi.org/10.1042/bj1490289
  3. Brendler-Schwaab, S., Czich, A., Epe, B., Gocke, E., Kaina, B., Muller, L., Pollet, D. and Utesch, D. (2004): Photochemical genotoxicity: principles and test methods. Report of a GUM task force. Mutat. Res., 566, 65-91 https://doi.org/10.1016/S1383-5742(03)00052-8
  4. Chetlat, A.-A., Albertini, S. and Gocke, E. (1996): The photomutagenicity of fluoroquinolones in tests for gene mutation, chromosomal aberration, gene conversion and DNA breakage (Comet assay). Mutagenesis, 11, 497-504 https://doi.org/10.1093/mutage/11.5.497
  5. Collins, A.R. (2004): The comet assay for DNA damage and repair: principles, applications, and limitations. Mol. Biotechnol., 26, 249-261 https://doi.org/10.1385/MB:26:3:249
  6. Davies, R.E. and Forbes, P.D. (1988): Retinoids and photocarcinogenesis. J. Toxicol.-Cutan. Ocul. Toxicol., 7, 241-253 https://doi.org/10.3109/15569528809056304
  7. Fu, P.P., Cheng, S.-H., Coop, L., Xia, Q., Culp, S.J., Tolleson, W.H., Wamer, W.G. and Howard, P.C. (2003): Photoreaction, Phototoxicity, and Photocarcinogenicity of Retinoids. J. Environ. Sci. Health Pt. C-Environ. Carcinog. Ecotoxicol. Rev., 21, 165-197 https://doi.org/10.1081/GNC-120026235
  8. Gocke, E., Muller, L., Guzzie, P.J., Brendler-Schwaab, S., Bulera, S., Chignell, C.F., Henderson, L.M., Jacobs, A., Murli, H., Snyder, R.D. and Tanaka, N. (2000): Consideration on photochemical genotoxicity: Report of the international workshop on genotoxicity test procedures working group. Environ. Mol. Mutagen., 35, 173-184 https://doi.org/10.1002/(SICI)1098-2280(2000)35:3<173::AID-EM4>3.0.CO;2-E
  9. Hong, M.-Y., Kim, J.-Y., Chung, M.K. and Lee, M. (2005): Prediction of photo-carcinogenicity from photo-Ames assay. Environ. Mutagens Carcinogens (Korean), 25, 6-12
  10. Jacobs, A., Avalos, J., Brown, P. and Wilkin, J. (1999): Does photosensitivity predict photococarcinogenicity? Int. J. Toxicol., 18, 191-198 https://doi.org/10.1080/109158199225486
  11. Jose, J.G. (1979): Photomutagenesis by chlorinated phenothiazine tranquilizers. Proc. Natl. Acad. Sci. U.S.A., 76, 469-471
  12. Kelly, G.E., Meikle, W.D. and Moore, D.E. (1989): Enhancement of UV-induced skin carcinogenesis by azathioprine: role of photochemical sensitization. Photochem. Photobiol., 49, 59-65 https://doi.org/10.1111/j.1751-1097.1989.tb04078.x
  13. Kelly-Garvert, F. and Legator, M.S. (1973): Photoactivation of chlorpromazine: cytogenetic and mutagenic effects. Mutat. Res., 21, 101-105 https://doi.org/10.1016/0165-1161(73)90005-8
  14. Meunier, J.-R., Sarasin, A. and Marrot, L. (2002): Photogenotoxicity of mammalian cells: A review of the different assays for in vitro testing. Photochem. Photobiol., 75, 437-447 https://doi.org/10.1562/0031-8655(2002)075<0437:POMCAR>2.0.CO;2
  15. Nagayo, K., Way, B.H., Tran, R.M. and Song, P.S. (1983): Photocarcinogenicity of 8-methoxypsoralen and aflatoxin B1 with longwave ultraviolet light. Cancer Lett., 18, 191-198 https://doi.org/10.1016/0304-3835(83)90067-8
  16. Olive, P.L., Banath, J.P. and Durand, R.E. (1990): Heterogeneity in radiation-induced DNA damage and repair in tumor and normal cells measured using the 'comet' assay. Radiat. Res., 122, 86-94 https://doi.org/10.2307/3577587
  17. Singh, N.P., McCoy, M.T., Tice, R.R. and Schneider, E.L. (1988): A simple technique for quantitation of low levels of DNA damage in individual cells. Exp. Cell Res., 175, 184-191 https://doi.org/10.1016/0014-4827(88)90265-0
  18. Speit, G. and Hartmann, A. (2006): The comet assay: a sensitive genotoxicity test for the detection of DNA damage and repair. Methods Mol. Biol., 314, 275-286 https://doi.org/10.1385/1-59259-973-7:275
  19. Spielmann, H., Muller, L., Averbeck, D., Balls, M., Brendler- Schwaab, S., Castell, J.V., Curren, R., de Silva, O., Gibbs, N.K., Liebsch, M., Lovell, W.W., Merk, H.F., Nash, J.F., Neumann, N.J., Pape, W.J.W., Ulrich, P. and Vohr, H.-W. (2000): The second ECVAM workshop on phototoxicity testing: The report and recommendations of ECVAM workshop 42. Altern. Lab. Anim., 28, 777-814
  20. Stern, R.S., Thibodeau, L.A., Kleinerman, R.A., Parrish, J.A. and Fitzpatrick, T.B. (1979): Risk of cutaneous carcinoma in patients treated with oral methoxsalen photochemotherapy for psoriasis. New Engl. J. Med., 300, 809-813 https://doi.org/10.1056/NEJM197904123001501
  21. Tice, R.R., Agurell, E., Anderson, D., Burlinson, B., Hartmann, A., Kobayashi, H., Miyamae, Y., Rojas, E., Ryu, J.C. and Sasaki, Y.F. (2000): Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ. Mol. Mutagen., 35, 206-221 https://doi.org/10.1002/(SICI)1098-2280(2000)35:3<206::AID-EM8>3.0.CO;2-J