Environmental Analysis Health and Toxicology

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

Evaluation of the Genetic Toxicity of Synthetic Chemicals (Ⅵ) -In vitro Chromosomal Aberration Assay with 17 Chemicals in Chinese Hamster Lung Cells -

합성화학물질들의 유전독성평가(Ⅵ) -Chinese hamster lung세포를 이용한 17종 합성화학물질들의 염색체 이 상 시험 -

  • Ryu, Jae-Chun (Toxicology Laboratory, Korea Institute of Science and Technology) ;
  • Kim, Kyung-Ran (Toxicology Laboratory, Korea institute of Science and Technology) ;
  • Kim, Youn-Jung (Toxicology Laboratory, Korea Institute of Science and Technology) ;
  • Jeon, Hee-Kyung (Toxicology Laboratory, Korea Institute of Science and Technology)
  • Published : 2003.06.01
Download PDF
⟨ Previous Next ⟩

Abstract

The validation of many synthetic chemicals that may pose a genetic hazard in our environment is of great concern at present. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of chemicals that possibly threaten the public health. In this respect, the regulation and evaluation of the chemical hazard playa very important role to environment and human health. The clastogenicity of 17 synthetic chemicals was evaluated in Chinese hamster lung fibroblast cells in vitro. 2-Nitroaniline (CAS No. 88-74-4) induced chromosomal aberrations with statistical significance at the concentration of 86.3 ${\mu}$g/ml in the absence of metabolic activation system. 1-Chloroanthraquinone (CAS No. 82-44-0) which is one of the most cytotoxic chemical among 17 chemicals tested revealed no clastogenicity in the range of 0.8 ∼ 3.0 ${\mu}$g/ml both in the presence and absence of S-9 metabolic activation system. From the results of chromosomal aberration assay with 17 synthetic chemicals in Chinese hamster lung cells in vitro, 2-Nitroaniline (CAS No. 88-74-4) revealed weak positive clastogenic results in this study.

합성화학물질들이 환경중에 유입되어 인체에는 물론 환경생태계에 많은 영향을 미치고 있어 이들의 유해성 검증은 매우 중요한 일이라 할 수 있다. 실제 산업체에서 사용되는 수많은 화학물질들의 유전적 손상 유발유무는 더욱이 중요한 일이라 할 수 있다. 이에 따라 산업체 공정과정에서 널리 사용되는 17종의 화학물질에 대해 Chinese hamster lung(CHL)세포를 이용한 염색체 이상 시험을 수행하여 이들이 염색체상에 구조적 이상을 유발하는지 관찰하였다. 그 결과, 본 연구에서 사용한 17종의 합성화학물질 중 2-nitroaniline(CAS No. 88-74-4)만이 대사활성화계 부재시 86.3 $\mu\textrm{g}$/ml의 농도에서 통계적으로 유의한 염색체 이상 유발능을 보였다. 반면 가장 높은 세포독성을 보인 1-chloroanthraquinone (CAS No. 82-44-0)은 0.8 ∼ 3.0 $\mu\textrm{g}$/ml의 시험농도범위에서 대사활성 존재 유무와 무관하게 염색체 이상을 유발하지 않았으며, 다른 15종의 물질들 역시 본시험 적용 농도 범위에서 염색체 이상 유발능을 관찰할 수 없었다.

Keywords

References

  1. Altman DG. Comparing groups-categorical data : Practical Statistics for Medical Research. Chapman & Hall, London, 1993; 229-276
  2. Ames BN, Durston WE, Yamasaki E and Lee FD. Carcinogens are mutagens : a simple test system combining liver homogenates for activation and bacteria for detection, Proc. Natl. Acad. Sci. USA 1973; 70: 2281-2285 https://doi.org/10.1073/pnas.70.8.2281
  3. Ames BN, McCann J and Yamasaki E. Method for detecting carcinogens and mutagens with Salmonella/mammalian-microsome mutagenicity test, Mutation Res 1975; 31: 347-364 https://doi.org/10.1016/0165-1161(75)90046-1
  4. Clive D, McCuen R, Spector JFS, Piper C and Mavournin KH. Specific gene mutations in L5178Y cells in culture: A report of the U.S. Environmental Protection Agency Gen-Tox Program, Mutation Res 1983; 115: 225-251 https://doi.org/10.1016/0165-1110(83)90005-2
  5. Galloway SM, Sofuni T, Shelby MD, Thilager A, Kumaroo V, Kaur P, Gulati D, Putman DL, Murli H, Marshall R, Tanaka N, Anderson B, Zeiger E and Ishidate M Jr. Multilaboratory comparison of in vitro tests for chromosome aberrations in CHO and CHL cells tested under the same protocols, Environ. Mol. Mutagenesis 1997; 29: 189-207 https://doi.org/10.1002/(SICI)1098-2280(1997)29:2<189::AID-EM10>3.0.CO;2-A
  6. Hayashi M, Sofuni T and Ishidate M Jr. High sensitivity in micronucleus induction of mouse strain, Mutation Res 1982; 105: 252-256
  7. Hayashi M, Morita T, Kodama Y, Sofuni T and Ishidate M Jr. The micronucleus assay with mouse peripheral blood reticulocytes using acridine orange-coated slides, Mutation Res 1990; 245: 245-249 https://doi.org/10.1016/0165-7992(90)90153-B
  8. Hayashi M, Kodama Y, Awogi T, Suzuki T, Asita AO and Sofuni T. The micronucleus assay using peripheral blood reticulocytes from mitomycin C- and cyclophosphamide-treated rats, Mutation Res 1992; 278: 209-213 https://doi.org/10.1016/0165-1218(92)90236-S
  9. Hayashi M, Maki-Paakkanen J, Tanabe H, Honma M, Suzuki T, Matsuoka A, Mizusawa H and Sofuni T. Isolation of micronuclei from mouse blood and fluorescence in situ hybridization with a mouse centromeric DNA probe, Mutation Res 1994; 307: 245-251 https://doi.org/10.1016/0027-5107(94)90298-4
  10. Henderson L, Jones E, Freemantle M, Howard CA, Jenkinson P, Lambert R, Mackay J, Marshall R and Wilcox P. Extended harvest times are not necessary for the detection of in vitro clastogens in regulatory cytogenetics studies, Mutagenesis 1996; 11: 61-67 https://doi.org/10.1093/mutage/11.1.61
  11. Heo MY, Kim JH and Ryu J-C. Anticlastogenicity of betacarotene and galangin using in vivo supravital staining micronucleus test, Environ. Mutagens & Carcinogens 1997; 17: 92-96
  12. Ishidate M and Odashima S. Chromosome test with 134 compounds on chinese hamster cells in vitro-A screening for chemical carcinogens, Mutation Res 1977; 48: 337-354 https://doi.org/10.1016/0027-5107(77)90177-4
  13. JEMS-MMS Atlas of chromosome aberration by chemicals, Japanese Environmental Mutagen Society-Mammalian Mutagenicity Study Group. Tokyo 1998
  14. Kim Y-J, Park H-J, Kim Y, Kim M-K, Lee S-H, Jung S-H and Ryu J-C. Genotoxicity Study of sophoricoside. A constituent of Sophora japonica, in bacterial and mammalian cell system, Environmental Mutagens & Carcinogens 2001; 21(2): 99-105
  15. Kohler SW, Provost GS, Fieck A, Kretz PL, Bullock WO, Sorge JA, Putman DL and Short JM. Spectra of spontaneous and mutagen-induced mutations in the lac I gene in transgenic mice, Proc Natl Acad Sci USA 1991; 88: 7958-7962 https://doi.org/10.1073/pnas.88.18.7958
  16. Maron DM and Ames BN. Revised methods for the Salmonella mutagenicity test, Mutation Res 1983; 113: 173-215 https://doi.org/10.1016/0165-1161(83)90010-9
  17. McCann J, Choi E, Yamasaki E and Ames BN. Detection of carcinogens as mutagens in the Salmonella/microsome test : assay of 300 chemicals, Proc Natl Acad Sci USA 1975; 72: 5135-5139 https://doi.org/10.1073/pnas.72.12.5135
  18. Mersch-Sundermann VS, Kerekordes S and Mochayedi S. Sources of variability of the Escherichia coli PQ 37 genotoxicity assay (SOS chromotest), Mutation Res 1991; 252: 51-60 https://doi.org/10.1016/0165-1161(91)90251-3
  19. Meselson M and Russell K. Comparison of carcinogenic and mutagenic potency. In Hiatt, H.H., Watson, J.D. and Winstend, J.A. (Eds.). Origin of Human Cancer. Cold Spring Harbor Laboratory, New York, 1991; pp. 1473-1481
  20. Mosmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays, J Immunol Methods 1983; 65: 55 -63 https://doi.org/10.1016/0022-1759(83)90303-4
  21. OECD. OECD giudeline for the testing of chemicals, Documents 473, Genetic toxicology: in vitro mammalian cytogenetic test, Organization for Economic Cooperation and Development, Paris, France. 1993
  22. Radman M, Jeggo P and Wagner R. Chromosomal rearrangement and carcinogenesis, Mutation Res 1982; 98: 249-264 https://doi.org/10.1016/0165-1110(82)90035-5
  23. Ryu J-C, Lee S, Kim K-R, Kim M, Chang I-M and Park J. A study on the clastogenicity of trichothecene mycotoxins in chinese hamster lung cells, Korean J Toxicol 1993a; 9: 13-21
  24. Ryu J-C, Kim S-H, Kim K-R, Lee S, Kim M, Jung H-K and Park J. A study on the antimutagenic effect of cinnamic acid derivatives in Escherichia coli PQ 37 (SOS Chromotest) (I), Environ Mutagens & Carcinogens 1993b; 13: 8-17
  25. Ryu J-C, Lee S, Kim K-R and Park J. Evaluation of the genetic toxicity of synthetic chemicals (I). Chromosomal aberration test on chinese hamster lung cells in vitro, Environ Mutagens & Carcinogens 1994; 14: 138 - 144
  26. Ryu J-C, Kim K-R, Kim H-J, Ryu E-K, Lee S-Y, Jung S-O, Youn J-Y, Kim M-H and Kwon O-S. Evaluation of the genetic toxicity of synthetic chemicals (II). a pyrethroid insecticide, fenpropathrin, Arch Pharm Res 1996a; 19: 251-257 https://doi.org/10.1007/BF02976235
  27. Ryu J-C, Kim K-R, Ryu E-K, Kim H-J, Kwon O-S, Song C-E, Mar W and Chang I-M. Chromosomal aberration assay of taxol and 10-deacetyl baccatin III in chinese hamster lung cells in vitro, Environ Mutagens & Carcinogens 1996b; 16: 6-12
  28. Ryu J-C, Kim H-J, Seo Y-R and Kim K-R. Single cell gel electrophoresis (comet assay) to detect DNA damage and apoptosis in cell level, Environ Mutagens & Carcinogens 1997; 17: 71-77
  29. Ryu J-C, Kim K-R, Kim H-J, Jung S-O, Kim M-K, Park H-S and Kim Y-H. Acute and Genetic Toxicity Study of DK 1002, a Drug Candidate for Analgesics, J Toxicol Pub Health 1998a; 14(3): 427-433
  30. Ryu J-C, Kim K-R, Kim H-J, Myung S-W, Kim G-H, Lee M-J and Chang I-M. Genotoxicity Study of Bojungchisup-tang, an oriental herbal decoction-in vitro chromosome aberration assay in chinese hamster lung cells and in vitro supravital-staining micronucleus assay with mouse peripheral reticulocytes, Arch Pharm Res 1998b; 21(4): 391-397 https://doi.org/10.1007/BF02974632
  31. Ryu J-C, Youn J-Y, Kim, Cho K-H and Chang I-M. Transgenic Mutagenesis assay to elucidate the mechanism of mutation in gene level, Environ Mutagen & Carcinogen 1998c; 18(1): 1-7 https://doi.org/10.1002/(SICI)1520-6866(1998)18:1<1::AID-TCM1>3.0.CO;2-K
  32. Ryu J-C, Youn J-Y, Cho K-H and Chang I-M. Mutation spectrum in lac I gene of transgenic Big Blue cell line following to short term exposure 4-nitroquinoline N-oxide, Environ Mol Mutagenesis 1998d; 31(29): p. 16
  33. Ryu J-C, Kim K-R and Choi Y-J. In vitro mouse lymphoma thymidine kinase (tk+/-) gene forward mutation assay in mammalian cells, Environ Mutagen & Carcinogen 1999a; 19(1): 7-13
  34. Ryu J-C, Youn J-Y, Kim Y-J, Kwon O-S, Kim Y-S, H-T, Cho K-H and Chang I-M. Mutation spectrum of 4-nitroquinoline N-oxide in the lac I transgenic Big Blue Rat2 cell line, Mutation Res 1999b; 445: 127-135 https://doi.org/10.1016/S1383-5718(99)00136-9
  35. Ryu J-C, Kim Y-J, Kim H-T and Chai Y-G. Genotoxicity Assessment of atrazine in the Big Blue rat2 lac I transgenic cell line, Environ Mol Mutagenesis 2000; 35(31): p. 52 (No. 176)
  36. Ryu J-C, Seo Y-R, Smith MA and Han SS. The Effect of methyl methanesulfonate (MMS)-induced excision repair on p53-dependent apoptosis in human lymphoid cells, Res Comm Mol Pathol and Pharmacol 2001 a; 109(1,2): 35-51
  37. Ryu J-C, Kim K-R, Lee S and Park J. Evaluation of the genetic toxicity of synthetic chemicals (III), Chromosomal aberration assay with 28 chemicals in chinese hamster lung cells in vitro, Environ Mutagens & Carcinogens 2001b; 21(1): 14-22
  38. Ryu J-C and Park KY. Anticlastogenic effect of Baechu (Chinese cabbage) Kimchi and Buchu (leek) Kimchi in supravital staining micronucleus assay using peripheral reticulocytes of mouse, Environ Mutagens & Carcinogens 2001c; 21(1): 51-56
  39. Ryu J-C, Kwon O-S and Kim H-T. Optimal conditions of Single Cell Gel Electrophoresis (Comet) Assay to detect DNA single strand breaks in mouse lymphoma L5178Y cells, Environ Mutagens & Carcinogens 2001d; 21(2): 89-94
  40. Ryu J-C, Kim Y-J and Chai Y-G. Mutation spectrum of 1, 2-dibromo-3-chloropropane, an endocrine disruptor, in the lac I tranasgenic Big Blue Rat2 fibroblast cell line, Mutagenesis 2002a; 17(4): 301-307 https://doi.org/10.1093/mutage/17.4.301
  41. Ryu J-C, Kim HT and Kim YJ. Studies on DNA single strand break of seven phthalate analogues in mouse lymphoma cells, Environ Mutagens & Carcinogens 2002b; 22(3): 164-168
  42. Ryu J-C, Kim KR and Kim YJ. Evaluation of the Genetic Toxicity of Synthetic Chemicals (IV) - in vitro chromosomal aberration assay with 18 chemicals in Chinese hamster lung cells-, Environ Mutagens & Carcinogens 2002c; 22(3): 149-156
  43. Ryu J-C, Kim KR, Kim YJ and Choi HY. Evaluation of the Genetic Toxicity of Synthetic Chemicals (V) - in vitro chromosomal aberration assay with 17 chemicals in Chinese hamster lung cells-, Environ Mutagens & Carcinogens, 2002d; 22(4): 215-222
  44. Ryu J-C. Current Trend of Advanced Methods in Cellular and Molecular Toxicology, Environ Mutagens & Carcinogens 2002e; 22(1): 1-11
  45. Ryu J-C. Introduction of Recent advanced cellular and Molecular Toxicology, Korean J Pesticide Science 2002f; 6(3): 157-165
  46. Sato T, Chikazawa K, Yamamori H, Ose Y, Nagase H. and Kito H. Evaluation of the SOS chromotest for the detection of antimutagens, Environ Mol Mutagenesis 1991; 17: 258-263 https://doi.org/10.1002/em.2850170407
  47. Sawyer J, Moore MM, Clive D and Hozier J. Cytogenetic characterization of the L5178Y TK+/- 3.7.2C mouse lymphoma cell line, Mutation Res 1985; 147: 243-253 https://doi.org/10.1016/0165-1161(85)90065-2
  48. Schmid W. The micronucleus test, Mutation Res 1975; 31: 9-15 https://doi.org/10.1016/0165-1161(75)90058-8
  49. Singh PN, Stephens RE and Schneider EL. Modification of alkaline microgel electrophoresis for sensitive detection of DNA damage, Int J Radiation Biol 1994; 66: 23-28 https://doi.org/10.1080/09553009414550911
  50. Suzuki T, Hayashi M, Sofuni T and Myhr BC. The concomitant detection of gene mutation and micronucleus induction by mitomycin C in vivo using lac Z transgenic mice, Mutation Res 1993; 285: 219-224 https://doi.org/10.1016/0027-5107(93)90109-S
  51. Tice RR, Agurell E, Anderson D, Burlinson B, Hartmann A, Kobayashi H, Miyamae Y, Rojas E, Ryu J-C and Sasaki YF. Single cell gel/comet assay: Guidelines for in vitro and in vivo genetic toxicology testing., Environ Mol Mutagenesis 2000; 35(3): 206-221 https://doi.org/10.1002/(SICI)1098-2280(2000)35:3<206::AID-EM8>3.0.CO;2-J
  52. WHO. The evaluation and testing of drugs for mutagenicity, principles and problems, WHO Tech Rep Ser 1971; No. 482
  53. Zimmermann FK. Induction of mitotic gene conversion by mutagens, Mutation Res 1971; 11: 327-337