Toxicological Research

Korean Society of Toxicology & Korea Environmental Mutagen Society (한국독성학회)
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Comparison of L5178Y tk+/- Mouse Lymphoma Assay and In vitro Chromosome Aberration Test

  • Lee, Michael (Korea Institute of Toxicology, Korea Research Institute of Chemical Technology) ;
  • Jung Kwon (Korea Institute of Toxicology, Korea Research Institute of Chemical Technology) ;
  • Cho, Ji-Hee (Korea Institute of Toxicology, Korea Research Institute of Chemical Technology) ;
  • Hong, Mi-Young (Korea Institute of Toxicology, Korea Research Institute of Chemical Technology) ;
  • Kim, Eun-Joo (Korea Institute of Toxicology, Korea Research Institute of Chemical Technology) ;
  • Junghee Han (Korea Institute of Toxicology, Korea Research Institute of Chemical Technology) ;
  • Chung, Moon-Koo (Korea Institute of Toxicology, Korea Research Institute of Chemical Technology) ;
  • Han, Sang-Seop (Korea Institute of Toxicology, Korea Research Institute of Chemical Technology)
  • Published : 2002.06.01
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Abstract

The mouse lymphoma assay (MLA) has been recently validated as a sensitive and specific test system to determine the genotoxic potential for a chemical. The objective of this study is to evaluate the utility of MLA for detecting mutagens. Especially, to compare MLA with the in vitro chromosomal aberration test (CA), we performed MLA using the microwell method with three chemicals (hydroxyurea, theophylline and amino acid copper complex), which were reportedly positive in the CA. In cell treated with hydroxyurea, anti-neoplastic agent that blocks DNA replication, evidence of a positive response was obtained without S9 mix for 4 h and 24 h. In addition, analysis of colony size distribution at concentration that gave an elevated mutant fraction showed that hydroxyurea induced a high proportion of small type colonies, indicating that hydroxyurea-induced mutation is associated with large chromosomal deletion. Conversely, negative MLA result was obtained for theophylline, which was wed as central nervous system stimulator. Although theophylline increased the mutant frequency at concentration of 1250 $\mu\textrm{g}$/$\textrm{m}{\ell}$ with S9 mix for 4 h, a concentration-related increase in mutant frequency was not observed. The MLA result of amino acid copper complex was considered equivocal because the positive result was obtained at concentration showing 10% or less RS or RTG. Thus, among 3 CA-positive chemicals, positive MLA result was obtained for one. The other two chemicals were negative and equivocal. However MLA, which evaluates mutagenic potential of chemicals through colony formation by cell grouth, may provide a higher predictivity of carcinogenesis than CA.

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References

  1. Mutat.Res. v.72 Point mutations at the thymidine kinase locus in L5178Y mouse lymphoma cells Amacher,D.E.;Paillet,S.C.;Turner,G.N.;Ray,V.A.;Salsburg,D.S.
  2. Cancer Lett. v.88 Induction of apoptosis in chronic myelogenous leukemia lymphocytes by hydroxyures and adriamycin Anand,S.;Verma,H.;Kumar,L.;Singh,N.
  3. Proc.Natl.Acad.Sci. v.87 Molecular dissection of mutations at the heterozygous thymidine kinase locus in mouse lymphoma cells Applegate,M.L.;Moore,M.M.;Broder,C.B.;Burrell,A.;Juhn,G.;Kasweck,K.L.;Lin,P.F.;Wadhams,A.;Hozier,J.C.
  4. Mutat.Res. v.224 Chromosome analysis of small and large L5178Y mouse lymphoma cell colonies. Comparison of trifluorothymidine-resistant and unselected cell colonies from mutagen-treated and control cultures Blazak,W.F.;Los,F.J.;Rudd,C.J.;Caspary,W.J.
  5. Mut.Res. v.455 The mouse lymphoma assay Clements,J.
  6. Mutat.Res. v.16 A mutational assay system using the thymidine kinase locus in mouse lymphoma cells Clive,D.;Flamm,W.G.;Machesko,M.R.;Bernheim,N.J.
  7. Mutat.Res. v.189 Guide for performing the mouse lymphoma assay for mammalian cell mutagencity Clive,D.;Caspary,W.;Kirby,P.E.;Krehl,R.;Moore,M.;Mayo,J.;Oberly,T.J.
  8. The detection of gene mutations in cultured mammalian cells in Mutagenicity Testing;A practical approach Cole,J.;Arlett,C.F.;Vaenitt,S.(ed.);Parry,J.M.(ed.)
  9. Mutagenesis v.1 The mutagenicity of sodium fluoride to L5178Y [wild-type and T$K^+/-$(3.7.2C)] mouse lymphoma cells Cole, J.;Muriel, W.J.;Bridges, B.A.
  10. Mutagenesis v.10 The use of L5178Y mouse lymphoma cells to assess the mutagenic, clastogenic and aneugenic properties of chemicals Combes,R.D.;Stopper,H.;Caspary,W.J.
  11. Mutat.Res. v.258 Consideration in the U.S. Environmental Protection Agency's testing approach for mutageneicity Dearfield,K.L.;Aueletta,A.E.;Clmino,M.C.;Moore,M.M.
  12. Ann.New York Acad.Sci v.76 Studies of the culture of leukemic cells in vitro Fischer,G.A.
  13. Environ.mol.Mutagen. v.25 International workshop on mouse lymphoma assay testing practices and data interpretations Garriot,M.L.;Casciano,D.A.;Schechtman,L.M.;Probst,G.S.
  14. Mutagenesis v.14 Evaluation of the mouse lymphoma tk assay (microwell method) as an alternative to the in vitro chromosomal aberration test Honma,M.;Hayashi,M.;Shimada,H.;Tanaka,N.;Wakuri,S.;Awogi,T.;Yamamoto,K.I.;Kodani,N.;Nishi,Y.;Nakadate,M.;Sofuni,T.
  15. Mutagenesis v.14 The need for long-term treatment in the mouse lymphoma assay Honma,M.;Zhang,L.Z.;Sakamoto,H.;Ozaki,M.;Takeshita,K.;Mosmose,M.;Hayashi,M.;Sofuni,T.
  16. Mutat.Res. v.84 Cytogenetic analysis of the L5178Y/T$K^+/-$ mouse lymphoma mutagenesis assay system Hozier,J.;Sawyer,J.;Moore,M.;Howard,B.;Clie,D.
  17. Step 4 Guideline Topics S2B Genotoxicity;A Standard Battery for Genotoxicity Testing for Pharmaceutical. International Conferences on the Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use ICH
  18. Data Book of Chromosome Aberration Test In Vitro Ishidata,M.Jr.
  19. Mutat.Res. v.415 Recommendations for spacing of test chemical concertrations in the mouse lymphoma tk mutation assay(MLA) Kirkland,D.J.;Clements,J.
  20. Mutagenesis v.13 Analysis of large and small colony L5178Y $tk^{+/-}$ mouse lymphoma mutants by loss heterozygosity (LOH) and by whole chromosome 11 painting;detection or recombination Liechty,M.C.;Scalzi,J.M.;Sims,K.R.;Crosby,Jr.H.;Spencer,D.L.;Davis,L.M.;Caspary,W.J.;Hozier,J.C.
  21. Mut.Res. v.394 The L5178Y/t$k^{+/-}$ mouse lymphoma specific gene and chromosomal mutation assay. A phase Ⅲ report of the U.S. enironmental protection agency gen-tox program Michell,A.D.;Auletta,A.E.;Clive,D.;Kirby,P.E.;Moore,M.M.;Myhr,B.C.
  22. Mutat.Res. v.151 Analysis of trifluorotymidine-resistant [TF$T^r$] mutants of L5178Y T$K^{+/-}$ mouse lymphoma cells Moore,M.M.;Clive,D.;Hozier,J.C.;Howard,B.E.;Batson,A.G.;Turner,N.T.;Sawyer,J.
  23. Environ.Mol.Mutat. v.35 Mouse lymphoma thymidine kinase locus gene mutation assay;International workshop on genotoxicity test procedures workgroup report Moore,M.M.;Honma,M.;Clements,J.;Awogi,T.;Bolcsfoldi,G.;Cole,J.;Gollapudi,B.;Harrington Brock,K.;Mitchell,A.;Muster,W.;Myhr,B.;O'Donovan,M.;Ouldelhkim,M.C.;San,R.;Shimada,H.;Stankowski,Jr.L.F.
  24. Mutat.Res. v.256 Recommended protocols based on a survey of current practice in genotoxicity testing laboratories;Ⅱ. Mutation in Chinese hamster ovay,V79 Chinese hamster lung and L5178Y mouse lymphoma cells Nestman,E.R.;Brillinger,R.L.;Gilman,J.P.W.;Rudd,C.J.;Swierenga,S.H.H.
  25. In vitro mammalian cell gene mutation test.Updated and adopted OECD Guideline 476
  26. J.Allergy Clin.Immunol. v.78 Overview of effects of theophylline Persson,C.G.
  27. Mutat.Res. v.257 International commission for protection against environmental mutagens and carcinogens Genotoxicity under extreme culture conditions[A report from ICPEMC Task Group 9.] Scott,D.;Galloway,S.M.;Marshall,R.R.;Ishidata,M.;Brusick,D.;Ashby,J.;Myhr,B.C.
  28. Mol.Cell.Biol. v.8 Effect of cycloheximide on development of methotrexate resistance of Chinese hamster ovary cells treated with inhibitors of DNA Synthesis Sherwood,S.W.;Schumacher,R.T.;Schmke.R.T.
  29. Environ.Mol.Mutat. v.29 Mouse lymphoma workshop;Victoria, British Columbia, Canada. Protocols issues regarding the use of the microwell method of the mouse lymphoma assay Sofuni,T.;Wilcox,P.;Shimada,H.;Clements,J.;Honma,M.;Clive,D.;Green,M.;Thybaud,V.;San,R.H.C.;Elliott,B.M.;Muller,L.
  30. Mutat.Res. v.488 A review of the genotoxicity of marketed pharmaceuticals Snyder,R.D.;Green,J.W.