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New ara-C resistant mutants of the Chinese hamster ovary cells

Published online by Cambridge University Press:  14 April 2009

N. C. Mishra ,
Kathryn Hinnant  and
Emily Cason
N. C. Mishra
Affiliation:
Department of Biology, University of South Carolina, Columbia, South Carolina 29208, U.S.A.
Kathryn Hinnant
Affiliation:
Department of Biology, University of South Carolina, Columbia, South Carolina 29208, U.S.A.
Emily Cason
Affiliation:
Department of Biology, University of South Carolina, Columbia, South Carolina 29208, U.S.A.
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Mutants of Chinese hamster ovary (CHO) cell resistant to cytosine arabinoside (ara-C), an inhibitor of DNA synthesis and antitumour drug, have been isolated and characterized both biochemically and genetically. Mutants occurring spontaneously and those induced by treatment with N-methyl-N′-Nitro-N-nitrosoguanidine (MNG), were obtained at a frequency of 0·24 × 10−6 and 3·4 10−6 respectively. The mutants showed a stable ara-C resistant phenotype which was inherited as a dominant trait in genetic crosses. The wild type (CHO K-1) and the mutant (103, 002 and 003) cells showed no differences in the levels of the uptake of ara-C or of its degradation. Results of biochemical studies further excluded the involvement of deaminase, kinase and ribonucleotide reductase as the possible factor(s) in conferring drug resistance to the mutant cells. However, the wild type and mutant DNA polymerases differed in the level of the in vitro incorporation of specific dNMP in the presence of ara-CTP. These data suggested that the wild-type DNA polymerase which becomes error prone in the presence of ara-CTP may cause the drug sensitivity of the wild-type cells and that a change in the mutant enzyme making it resistant (or less prone) to ara-CTP induced errors in dNMP incorporation may control the drug resistance of the mutant cells.

Type
Research Article
Information
Genetics Research , Volume 45 , Issue 3 , June 1985 , pp. 265 - 277
Copyright
Copyright © Cambridge University Press 1985

References

REFERENCES

Boehn, T. L. J. & Drahorsky, D. (1982). Elevated level of enzymatic DNA methylation in cells treated with ara-C. Cancer Research 42, 1537–1540.Google Scholar
Bell, D. E. & Fridland, A. (1980). Mode of action of 9-β-D-arabinosyladenine and 2-β- D-arabinosylcytosine on DNA synthesis in human lymphoblasts. Biochimia et biophysica acta 606, 57–66.CrossRefGoogle Scholar
Cohen, S. C. (1977). The mechanisms of lethal action of arabinosyl cytosine (ara-C) and arabinosyl adenine (ara-A). Cancer 40, 509–518.3.0.CO;2-8>CrossRefGoogle ScholarPubMed
Cohen, S. C. & Plunkett, W. (1975). Utilization of nucleosides by animal cells. Annals of the New York Academy of Science 255, 269–286.CrossRefGoogle ScholarPubMed
Chu, M. Y. & Fisher, G. A. (1965). A comparative study of leukemia cells sensitive and resistant to ara-C. Biochemical Pharmacology 14, 333–341.CrossRefGoogle Scholar
Chu, M. Y. & Fischer, G. A. (1965). Comparative studies of leukemic cells sensitive and resistant to cytosine arabinoside. Biochemical Pharmacology 14, 333–341.CrossRefGoogle ScholarPubMed
Davidson, R. L. & Gerald, S. (1976). Improved techniques for induction of mammalian cell hybridization by polyethylene glycol. Somatic Cell Genetics 2, 165–176.CrossRefGoogle ScholarPubMed
Eagle, H. (1959). Amino acid metabolism in mammalian cell cultures. Science 136, 432–437.CrossRefGoogle Scholar
Harris, M. (1974). Comparative frequency of dominant and recessive markers for drug resistance. Journal of the National Cancer Institute 52, 1811–1816.CrossRefGoogle ScholarPubMed
Harris, M. (1982). Induction of thymidine kinase in enzyme deficient Chinese hamster cell. Cell 25, 483–492.CrossRefGoogle Scholar
Hirsch, M. S. & Schooley, R. T. (1983). Drug therapy. New England Journal of Medicine 309, 963–970.Google ScholarPubMed
Huberman, E. & Heidelberger, C. (1982). The mutagenicity to mammalian cells of pyrimidine nucleoside analogs. Mutation Research 14, 130–132.CrossRefGoogle Scholar
Jacoby, W. B. & Pastan, I. M. (1970). Cell culture. Methods in Enzymology 58, 3–221.Google Scholar
Ives, D. & Wang, S. M. (1971). Deoxycytidine kinase from calf thymus. Methods in Enzymology 51, 337–345.CrossRefGoogle Scholar
Hopfee, P. A., Hunt, S. W. & Chiang, J. (1982). Isolation of deoxycoformycin-resistant cells with increased levels of adenosine deaminase. Somatic Cell Genetics 8, 465–477.CrossRefGoogle Scholar
Kessel, D. & Sharin, S. B. (1968). Transport of two nonmetabolized nucleosides deoxycytidine and cytosine arabinoside in a subline L1210 murine leukemia. Biochemical et biophysical acta 163, 179–187.CrossRefGoogle Scholar
Peterson, W. D., Simpson, W. F. & Hukku, B. (1979). Cell culture characterization: monitoring for cell identification. Methods in Enzymology 58, 166–178.Google ScholarPubMed
Plunkett, W., Lapi, L., Ortiz, P. J. & Cohen, S. (1974). Penetrance of mouse fibroblasts by 5′ phosphate of β-D-arabinofuranosyladenine and incorporation of the nucleotide into DNA. Proceedings of the National Academy of Sciences of the USA 71, 73–77.CrossRefGoogle Scholar
Puck, T. T. (1972). The Mammalian Cell as a Micro-organism, pp. 1–230. San Francisco: Holden Day.Google Scholar
Reddy, G. P. & Pardee, A. B. (1980). Multi-enzyme complex for metabolic channeling in mammalian DNA replication. Proceedings of the National Academy of Sciences of the USA 77, 3312–3316.CrossRefGoogle Scholar
Robert De Saint, B. & Buttin, G. (1979). Studies on 1β-D-arabinofuranosyl cytosine resistant mutants of chinese hamster fibroblast. Somatic Cell Genetics 5, 69–82.Google Scholar
Schrecker, A. W. & Urshel, M. J. (1968). Metabolism of ara-C in leukemia L1210 studies with intact cells. Cancer Research 28, 793–801.Google Scholar
Smith, D. B. & Chu, E. H. (1972). Genetic approach to the study of cytotoxicity and resistance of cultured Chinese hamster cells. Cancer Research 32, 1651–1657.Google Scholar
Steuart, C. D. & Burke, P. J. (1971). Cytidine deaminase and the development of resistance to arabinosyl cytosine. Nature New Biology 233, 109–110.CrossRefGoogle ScholarPubMed
Topal, M. D. & Baker, M. S. (1982). DNA precursor pool: A significant target for N-methyl-N-nitrosourea in C3H/10T ½ clone of cells. Proceedings of the National Academy of Sciences of the USA 79, 2211–2215.CrossRefGoogle Scholar
Wright, W. E. (1982). The selection of heterokaryons and cell hybrids using the biochemical inhibitors iodoacetamide and diethyl pyrocarbonate. In Techniques in Somatic Cell Genetics (ed. Shay, J. W.), pp. 47–65. New York: Plenum.CrossRefGoogle Scholar