Environmental Analysis Health and Toxicology

The Korean Society of Environmental Health and Toxicology (환경독성보건학회)
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Evaluation of the Genetic Toxicity of Synthetic Chemicals (Ⅷ) - In vivo Bone Marrow Micronucleus Assay of 8 Synthetic Chemicals in Mice -

합성화학물질들의 유전독성평가(Ⅷ) -마우스의 골수세포를 이용한 8종 합성화학물질들의 생체내 소핵시험-

  • 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)
  • Published : 2003.06.01
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Abstract

To validate and to estimate the chemical hazard playa very important role to environment and human health. The detection of many synthetic chemicals used in industry 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 resepct, the clastogenicity of 8 synthetic chemicals was evaluated with bone marrow micronucleus assay in mice. The positive control, mitomycin C(2mg/kg,i.p.) revealed significant induction ratio of percentage of micronucleated polychromatic erythrocytes/l,000 polychromatic erythrocytes compared to carboxymethylcellulose control. The chemicals with relatively high LD$\_$50/ value such as phenylisocyanate (CAS No. 103-71-9), m-aminochlorobenzene (CAS No. 108-42-9) and 2-chloro-4-nitroaniline (CAS No. 121-87-9) revealed no significant induction of micronucleated polychromatic erythrocytes in mice. From this results, 8 synthetic chemicals widely used in industry have revealed no significant micronucleus induction of clastogenicity in mice in this experiment.

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 산업체 공정과정에서 널리 사용되는 것으로 알려진 8종의 합성화학물질에 대해 마우스의 골수세포를 이용한 in vivo 소핵시험을 수행하여, 소핵형성 유발유무를 관찰하였다. 양성대조군으로 사용된 mitomycin는 음성대조군과 비교시 유의하게 소핵을 유발하는 반면, 비교적 마우스에서 높은 50% 치사량을 보이는 phenylisocyanate, m-aminochlorobenzene 및 2-chloro-4-nitroaniline 등의 합성물질들을 비롯한 나머지 5종의 물질들은 본 실험결과 통계적으로 유의하게 소핵을 유발하지 않는 것을 관찰 할 수 있었다.

Keywords

References

  1. 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
  2. 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
  3. Becker P, Scherthan H and Zankl H. Use of a centromerespecific DNA probe (p82H) in nonisotopic in situ hybridization for classification of micronuclei, Genes Chromosomes Cancer 1990; 2: 59-62 https://doi.org/10.1002/gcc.2870020111
  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. Clive D, Bolcsfoldi G, Clements J, Cole J, Homna Masamitsu, Majeska J, Moore M, Muller L, Myhr B, Oberly T, Oudelhkim M-C, Rudd C, Shimada H, Sofuni T, Thybaud V and Wilcox P. Consensus agreement regarding protocol issues discussed during the mouse lymphoma workshop, Environ. Mol. Mutagenesis 1995; 25: 165-168 https://doi.org/10.1002/em.2850250211
  6. Garriott ML, Casciano DA, Schechtman LM and Probst GS. International workshop on mouse lymphoma assay testing practices and data interpretations, Environ. Mol. Mutagenesis 1995; 25: 162-164 https://doi.org/10.1002/em.2850250210
  7. Hayashi M, Sofuni T and Ishidate M Jr. High sensitivity in micronucleus induction of mouse strain, Mutation Res 1982; 105: 252-256
  8. Hayashi M, Sofuni T and Ishidate M Jr. An application of acridine orange fluorescent staining to the micronucleus test, Mutation Res 1983; 120: 241-247 https://doi.org/10.1016/0165-7992(83)90096-9
  9. 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
  10. Hayashi M, Kodama Y, Awog 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
  11. 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
  12. Hayashi M, Ueda T, Uyeno K, Wada K, Kinae N, Saotome K, Tanaka N, akai A, Sasaki YF, Asano N, Sofuni T and Ojima Y. Development of genotoxicity assay systems that use aquatic organisms, Mutation Res 1998; 399: 125-133 https://doi.org/10.1016/S0027-5107(97)00251-0
  13. Heddle JA. A rapid in vivo test for chromosomal damage, Mutation Res 1973; 18: 187-190 https://doi.org/10.1016/0027-5107(73)90035-3
  14. 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
  15. IARC (International Agency for Research on Cancer). IARC monographs on the evaluation of carcinogenic risks to humans, Supplement 7, Overall Evaluation of Carcinogenicity: An updating of IARC monographs from Vols. 1 to 42, IARC, Lyon, 1987
  16. 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
  17. Komae N, Hibino Y and Sugano N. Analysis of micronuclei induced under hyperthermic conditions in human lymphocyte culture by fluorescence in situ hybridization (FISH) and spectral karyotyping (SKY) methods, Yakugaku Zasshi 1999; 119: 763-772 https://doi.org/10.1248/yakushi1947.119.10_763
  18. Lorke D. A new approach to practical acute toxicity testing, Arch Toxicol 1983; 54: 275-287 https://doi.org/10.1007/BF01234480
  19. 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
  20. MacGregor JT, Wehr CM and Gould DH. Clastogeninduced micronuclei in peripheral blood erythrocytes: The basis of an improved micronucleus test, Environ. Mutagen 1980; 2: 509-514 https://doi.org/10.1002/em.2860020408
  21. MacGregor JT, Wehr CM, Henika PR, Shelby MD. The in vivo erythrocyte micronucleus test: measurement at steady state increases assay efficiency and permits integration with toxicity studies, Fundam Appl Toxicol 1990; 14(3): 513-22 https://doi.org/10.1016/0272-0590(90)90255-I
  22. 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
  23. Meselson M and Russell K. Comparison of carcinogenesis and mutagenesis potency. Origin of Human Cancer edited by Hiatt, H.H., Watson, J.D. and Winstend, J.A. Cold Spring Harbor Laboratory, New York, 1991; 1473-1481
  24. Miller BM, Zitzelsberger HF, Weier H-Ul G and Adler I-D. Classification of micronuclei in murine erythrocytes: Immunofluorescent staining using CREST antibodies compared to in situ hybridization with biotinylated gamma satellite DNA, Mutagenesis 1991; 6: 297-302 https://doi.org/10.1093/mutage/6.4.297
  25. Morita T, Asano N, Awogi T, Sasaki Y F, Sato S, Shimada H, Sutou S, Suzuki T, Wakata A, Sofuni T and Hayashi M. Evaluation of the rodent micronucleus assay in the screening of IARC carcinogens (groups 1, 2A and 2B): The summary report of the 6th collaborative study by CSGMT/JEMS.MMS, Mutation Res 1997; 389: 3-122 https://doi.org/10.1016/S1383-5718(96)00070-8
  26. Oberly TJ and Garriott ML. Influence of cytotoxicity on test results in the L5178Y TK+/- mouse lymphoma assay, Environ. Mol. Mutagenesis 1996; 27: 75-78 https://doi.org/10.1002/(SICI)1098-2280(1996)27:1<75::AID-EM10>3.0.CO;2-G
  27. Peace BE and Succop P. Spontaneous micronucleus frequency and age: What are normal values? Mutation Res 1999; 425: 225-230 https://doi.org/10.1016/S0027-5107(99)00030-5
  28. 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 1993; 9: 13-21
  29. 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
  30. 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 1996a; 16: 6-12
  31. 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 1996b; 19: 251-257 https://doi.org/10.1007/BF02976235
  32. 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 1998; 21(4): 391-397 https://doi.org/10.1007/BF02974632
  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 1999; 19(1): 7-13
  34. Ryu J-C, Kim K-R, Lee S and Park J. Evaluation of the genetic toxicity of synthetic chemicals (Ill), Chromosomal aberration assay with 28 chemicals in chinese hamster lung cells in vitro, Environ Mutagens & Carcinogens 2001a; 21(1): 14-22
  35. 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 2001b; 21(1): 51-56
  36. 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 2002a; 22(3): 149-156
  37. 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. 2002b; 22(4): 215-222
  38. Saotome K, Sofuni T and Hayashi M. A micronucleus assay in sea urchin embryos, Mutation Res 1999; 446: 121-127 https://doi.org/10.1016/S1383-5718(99)00155-2
  39. 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
  40. Schmid, W. The micronucleus test, Mutation Res 1975; 31: 9-15 https://doi.org/10.1016/0165-1161(75)90058-8
  41. Tanisho T, Yagi T, Iwasaki K, Shimoi K, Kinae N, Hayashi M and Sofuni T. Monitoring of coastal water contaminated with mutagens and/ or carcinogens using micronucleus test in fish, Environ. Mutagen Res 1998; 20: 1-9
  42. Tice RR, Erexson GL, Hilliard CJ, Huston JL, Boehm RM, Gulati D, Shelby MD. Effect of treatment protocol and sample time on the frequencies of micronucleated polychromatic erythrocytes in mouse bone marrow and peripheral blood, Mutagenesis 1990; 5(4): 313-21 https://doi.org/10.1093/mutage/5.4.313
  43. Yamamoto KI and Kikuchi Y. A comparison of diameters of micronuclei induced by clastogens and by spindle poisons, Mutation Res 1980; 71: 127-131 https://doi.org/10.1016/0027-5107(80)90012-3