Abstract
The CHO/HGPRT system has been developed and defined for quantifying mutation induced by various physical and chemical agents at the hypoxanthine-guanine phosphoribosyl transferase (HGPRT) locus in Chinese hamster ovary (CHO) cells. Various genetic, biochemical, and physiological evidence supports the genetic basis of mutation induction in this system.
In all direct-acting chemical mutagens studies, mutation induction increased linearly as a function of the concentration, with no apparent threshold. Some chemicals induce mutation at non-cytotoxic concentrations; others induce mutation only with a concomitant loss of cell survival. In one dosimetry study, the mutagenicity of ethyl methanesulfonate has been quantified as a function of exposure concentration × treatment time. The sensitive and quantitative nature of the system enables studies of the structure-activity (mutagenicity) relationships of various classes of chemicals, including alkylating agents, heterocyclic nitrogen mustards, and platinum compounds. When rat liver S-9-mediated metabolic activation is present, procarcinogens such as benzo(a)-pyrene, 2-acetylaminofluorene, and dime thylnitrosamine are mutagenic, whereas their noncarcinogenic structural analogues pyrene, fluorene, and dimethylamine are not. Mutagenicity as determined in the assay appears to correlate well (77/84 = 92 percent) with the reported carcinogenicity in animals of 132 chemicals being examined. Quantification of mutagenicity of procarcinogens is complicated by the different optimum activation conditions required for different compounds such as benzo(a)pyrene and dimethylnitrosamine.
The system has been shown to be useful in determining the interactive effects between physical and chemical agents, and in screening for mutagenicity of a fractionated organic mixture derived from a liquified coal sample, and of industrial chemicals such as vinyl chloride in both liquid and gaseous state. For the system to be used successfully in routine screening, further studies should be directed toward the development of a metabolic activation system suitable for a broad spectrum of industrial and environmental chemicals, a sensitive and reliable statistical method well defined in consideration of specific intrinsic characteristics of the CHO/HGPRT protocol to clearly differentiate mutagenicity from nonmutagenicity, and an experimental design to determine compounds with low, yet detectable, mutagenicity.
The system has been expanded for determination of mutageninduced chromosome aberration, sister-chromatid exchange, and micronucleus formation in addition to gene mutation and cytotoxi city; it can also be used to study inhibition of DNA synthesis. Development of this Multiplex CHO Genetic Toxicology System should allow simultaneous determination of multiple, distinct biological end points and studies of interrelationships among these effects.
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Postdoctoral Fellows supported by the Monsanto Toxicology Fund.
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References
Barnhart, B. J., S. H. Cox, and R. T. Okinaka, 1978. Mutagenicity and cytotoxicity of light-activated procarcinogens. J. Cell Biol., 79: 388a.
Barskey, F. C., J. D. Irr, and D. F. Krahn, 1979. Mutagenicity of gases in Chinese hamster ovary cells assay. Proc. Ann. Meet. Environ. Mutagen Soc., p. 89.
Chu, E. H. Y., and S. S. Powell, 1976. Selective systems in somatic cell genetics. Adv. Hum. Genet., 7: 189–258.
Couch, D. B., and A. W. Hsie, 1976. Dose-response relationships of cytotoxicity and mutagenicity of monofunctional alkylating agents in Chinese hamster ovary cells. Mutat. Res., 38: 399.
Couch, D. B., and A. W. Hsie, 1978a. Metal mutagenesis: Studies of the mutagenicity of manganous chloride and 14 other metallic compounds in the CHO/HGPRT assay. Proc. Ann. Meet. Environ. Mutagen Soc., p. 74.
Couch, D. B., and A. W. Hsie, 1978b. Mutagenicity and cytotoxicity of congeners of two classes of nitroso compounds in Chinese hamster ovary cells. Mutat. Res., 57: 209–216.
Couch, D. B., N. L. Forbes, and A. W. Hsie, 1978. Comparative mutagenicity of alkylsulfate and alkanesulfonate derivatives in Chinese hamster ovary cells. Mutat. Res., 57: 217–224.
Deluca, J. G., D. A. Kaden, J. Krolewski, T. R. Shopek, and W. S. Thilly, 1977. Comparative mutagenicity of ICR-191 to S. typhimurium and diploid human lymphoblasts. Mutat. Res., 46: 11–18
Fuscoe, J. C., J. P. O’Neill, and A. W. Hsie, 1979. Structure-activity relationship of 19 antitumor ICR compounds in the CHO/HGPRT system. Proc. Ann. Meet. Environ. Mutagen Soc., p. 112.
Hsie, A. W., P. A. Brimer, T. J. Mitchell, and D. G. Gosslee, 1975a. The dose-response relationship for ethyl methanesul-fonate-induced mutations at the hypoxanthine-guanine phosphoribosyl transferase locus in Chinese hamster ovary cells. Somat. Cell Genet., 1: 247–261.
Hsie, A. W., P. A. Brimer, T. J. Mitchell, and D. G. Gosslee, 1975b. The dose-response relationship for ultraviolet light- induced mutations at the hypoxanthine-guanine phosphoribosyl transferase locus in Chinese hamster ovary cells. Somat. Cell Genet., 1: 383–389.
Hsie, A. W., A. P. Li, and R. Machanoff, 1977a. A fluence-response study of lethality and mutagenicity of white, black, and blue fluorescent light, sunlamp, and sunlight irradiation in Chinese hamster ovary cells. Mutat. Res., 45 333–342.
Hsie, A. W., P. A. Brimer, R. Machanoff, and M. H. Hsie, 1977b. Further evidence for the genetic origin of mutations in mammalian somatic cells: The effects of ploidy level and selection stringency on dose-dependent chemical mutagenesis to purine analogue resistance in Chinese hamster ovary cells. Mutat. Res., 45: 271–282.
Hsie, A. W., R. Machanoff, D. B. Couch, and J. M. Holland, 1978a. Mutagenicity of dimethyl nitrosamine and ethyl methanesul- fonate as determined by a quantitative host-mediated CHO/HGPRT assay, Mutat. Res., 51: 77–84.
Hsie, A. W., D. B. Couch, J. P. O’Neill, J. R. San Sebastian, P. A. Brimer, R. Machanoff, J. C. Riddle, A. P. Li, J. C. Fuscoe, N. L. Forbes, and M. H. Hsie, 1978b. Utilization of quantitative mammalian cell mutation system, CHO/HGPRT, in experimental mutagenesis and genetic toxicology, pp. 39–54. In: “Strategies for Short-Term Testing for Mutagens/Carcinogens,” B. E. Butterworth (ed.). CRC Press, Inc., West Palm Beach, Florida. Also in Proceedings of Chemical Industry Institute of Toxicology (CUT) Workshop on “Strategies for Short-term Testing for Mutagens/Carcinogens,” 1977. Research Triangle Park, North Carolina.
Hsie, A. W., J. P. O’Neill, D. B. Couch, J. R. San Sebastian, P. A. Brimer, R. Machanoff, J. C. Fuscoe, J. C. Riddle, A. P. Li, N. L. Forbes, and M. H. Hsie, 1978c. Quantitative analyses of radiation-and chemical-induced lethality and mutagenesis in Chinese hamster ovary cells. Radiat. Res., 76: 471–492.
Hsie, A. W., J. P. O’Neill, J. R. San Sebastian, D. B. Couch, P. A. Brimer, W. N. C. Sun, J. C. Fuscoe, N. L. Forbes, R. Machanoff, J. C. Riddle, and M. H. Hsie, 1978d. Quantitative mammalian cell genetic toxicology: Study of the cytotoxicity and mutagenicity of 70 individual environmental agents related to energy technologies and three subfractions of a crude synthetic oil in the CHO/HGPRT system.In: “Symposium on Short-Term Bioassays in the Fractionation and Analysis of Complex Environmental Mixtures.” Williamsburg, Virginia.
IARC, 1972–1976. “Monograph on the Evaluation of Carcinogenic Risk of Chemicals to Man,” Vols. 1–10. International Agency for Research on Cancer, Lyon, France.
Johnson, N. P., J. D. Hoeschele, R. O. Rahn, and A. W. Hsie, 1979. Mutagencity, cytotoxicity, and DNA-binding of Pt(II)chloroamines in CHO cells. Proc. Ann. Meet. Environ. Mutagen Soc., p. 85.
McCann, J., E. Choi, E. Yamasaki, and B. N. Ames, 1975. Detection of carcinogens as mutagens in the Salmonella/microsome test: Assay of 300 chemicals. Proc. Natl. Acad. Sci. (USA), 72: 5135–5139.
O’Neill, J. P., and A. W. Hsie, 1977. Chemical mutagenesis of mammalian cells can be quantified. Nature, 269:815–817.
O’Neill, J. P., P. A. Brimer, R. Machanoff, G. P. Hirsch, and A. W. Hsie, 1977a. A quantitative assay of mutation induction at the hypoxanthine-guanine phosphoribosyl transferase locus in Chinese hamster ovary cells: Development and definition of the system. Mutat. Res., 45: 91–101.
O’Neill, J. P., P. A. Brimer, R. Machanoff, J. R. San Sebastian, P. A. Brimer, and A. W. Hsie, 1977b. A quantitative assay of mutation induction at the hypoxanthine-guanine phosphoribosyl transferase locus in Chinese hamster ovary cells (CHO/HGPRT system): Utilization with a variety of mutagenic agents. Mutat. Res., 45: 103–109.
O’Neill, J. P., J. C. Fuscoe, and A. W. Hsie, 1978a. Mutagenicity of heterocyclic nitrogen mustards (ICR compounds) in cultured mammalian cells. Cancer Res., 38:506–509.
O’Neill, J. P., J. C. Fuscoe, and A. W. Hsie, 1978b. Structure-activity relationship of antitumor agents in the CHO/ HGPRT
O’Neill, J. P., and A. W. Hsie, 1980. The CHO/HGPRT mutagenicity assay. II: Genetic basis of 6-thioguanine resistance. In: “Quantitative Mammalian Cell Mutagenesis and Mutagen Screening,” A. W. Hsie, J. P. O’Neill, and V. K. McElheny (eds.). Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
O’Neill, J. P., and A. W. Hsie, 1979. Phenotypic expression time of mutagen-induced 6-thioguanine resistance in Chinese hamster ovary cells (CHO/HGPRT system). Mutat. Res., 59 109–118.
O’Neill, J. P., R. Machanoff, and A. W. Hsie, 1979. The use of a rat liver S-9 activation system in the CHO/HGPRT mutation induction assay. Proc. Ann. Meet. Environ. Mutagen Soc., p. 67.
Painter, R. B., 1977. Rapid test to detect agents that damage human DNA. Nature, 265:650–651.
Peck, P. M., T. K. Tan, and E. B. Peck, 1976a. Pulmonary carcinogenesis by derivatives of polynuclear aromatic alkylating agents. Cancer Res.,36: 2423–2427.
Peck, P. M., T. K. Tan, and E. B. Peck, 1976b. Carcinogenicity of derivatives of polynuclear compounds. J. Med. Chem., 19: 1422–1423.
Puck, T. T., 1972. “The Mammalian Cell as a Microorganism.” Holden-Day, Inc., San Francisco. 219 p.
Riddle, J. C., and A. W. Hsie, 1978. An effect of cell cycle position on ultraviolet-light-induced mutagenesis in Chinese hamster ovary cells. Mutat. Res., 52: 409–420.
San Sebastian, J. R., J. P. O’Neill, and A. W. Hsie, 1979. A multiplex CHO genetic toxicology system: Simultaneous determination of cytotoxicity, mutation induction, chromosome aberration, and sister chromatid exchange in mammalian cells. Proc. Ann. Meet. Environ. Mutagen Soc., p. 111.
Schmid, W., 1975. The micronucleus test. Mutat. Res., 31 9–15.
Siminovitch, L., 1976. On the nature of heritable variation in cultured somatic cells. Cell, 7:1–11.
Sirover, M. A., and L. A. Loeb, 1976. Metal-induced infidelity during DNA synthesis. Proc. Natl. Acad. Sci. (USA), 73: 2331–2335.
Thielmann, H. W., C. H. Schroder, P. A. Brimer, A. W. Hsie, and J. P. O’Neill, 1979. Relationship between DNA alkylation and specific-locus mutation induction by N-methyl- and N-ethyl- N-nitrosourea in cultured Chinese hamster ovary cells (CHO/ HGPRT system). Chem. Biol. Interact., 26(3): 233–243.
Thompson, L. H., and R. M. Baker, 1973. Isolation of mutants of cultured mammalian cells. Methods Cell Biol., 6:209–281.
USPHS, 1973–1976. “Survey of Compounds Which Have Been Tested for Carcinogenic Activity.” United States Public Health Service. USPHS Publication No. 149.
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Hsie, A.W., O’Neill, J.P., San Sebastian, J.R., Brimer, P.A., Tan, EL. (1983). Quantitative Mammalian Cell Mutagenesis and Mutagen Screening: Study with Chinese Hamster Ovary Cells. In: Kolber, A.R., Wong, T.K., Grant, L.D., DeWoskin, R.S., Hughes, T.J. (eds) In Vitro Toxicity Testing of Environmental Agents. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-3566-5_13
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