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. 1986 Aug;83(16):5978–5982. doi: 10.1073/pnas.83.16.5978

Transient hypoxia enhances the frequency of dihydrofolate reductase gene amplification in Chinese hamster ovary cells.

G C Rice, C Hoy, R T Schimke
PMCID: PMC386420  PMID: 3461470

Abstract

Exposure of Chinese hamster cells to reduced oxygen partial pressure results in a marked enhancement in the frequency of methotrexate resistance and dihydrofolate reductase gene amplification. The frequency of enhanced resistance is a function of the length of exposure to hypoxic conditions and the time after recovery from hypoxia when cells are plated into methotrexate-containing medium. Hypoxia results in an inhibition of DNA synthesis; upon return to normal oxygen atmosphere, greater than 60% of cells in S phase at the time hypoxia was started subsequently undergo overreplication of DNA within a single cell cycle. The cells with the increased frequency of gene amplification are derived from this subset of overreplicated cells. These results are discussed within the context of the hypoxic state of many solid tumors and the high frequency of aneuploidy, chromosomal aberrations, and spontaneously occurring resistances to a number of cancer chemotherapeutic agents.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Billen D. Replication of the bacterial chromosome: location of new initiation sites after irradiation. J Bacteriol. 1969 Mar;97(3):1169–1175. doi: 10.1128/jb.97.3.1169-1175.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brown P. C., Tlsty T. D., Schimke R. T. Enhancement of methotrexate resistance and dihydrofolate reductase gene amplification by treatment of mouse 3T6 cells with hydroxyurea. Mol Cell Biol. 1983 Jun;3(6):1097–1107. doi: 10.1128/mcb.3.6.1097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dolbeare F., Gratzner H., Pallavicini M. G., Gray J. W. Flow cytometric measurement of total DNA content and incorporated bromodeoxyuridine. Proc Natl Acad Sci U S A. 1983 Sep;80(18):5573–5577. doi: 10.1073/pnas.80.18.5573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Eberle H., Forrest N., Hrynyszyn J., Van Knapp J. Regulation of DNA synthesis and capacity for initiation in DNA temperature sensitive mutants of Escherichia coli I. Reinitiation and chain elongation. Mol Gen Genet. 1982;186(1):57–65. doi: 10.1007/BF00422912. [DOI] [PubMed] [Google Scholar]
  5. Frankfurt O. S., Slocum H. K., Rustum Y. M., Arbuck S. G., Pavelic Z. P., Petrelli N., Huben R. P., Pontes E. J., Greco W. R. Flow cytometric analysis of DNA aneuploidy in primary and metastatic human solid tumors. Cytometry. 1984 Jan;5(1):71–80. doi: 10.1002/cyto.990050111. [DOI] [PubMed] [Google Scholar]
  6. GOLDACRE R. J., SYLVEN B. On the access of blood-borne dyes to various tumour regions. Br J Cancer. 1962 Jun;16:306–322. doi: 10.1038/bjc.1962.36. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Haas M., Yoshikawa H. Defective bacteriophage PBSH in Bacillus subtilis. II. Intracellular development of the induced prophage. J Virol. 1969 Feb;3(2):248–260. doi: 10.1128/jvi.3.2.248-260.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hill A. B., Schimke R. T. Increased gene amplification in L5178Y mouse lymphoma cells with hydroxyurea-induced chromosomal aberrations. Cancer Res. 1985 Oct;45(10):5050–5057. [PubMed] [Google Scholar]
  9. Lavi S. Carcinogen-mediated amplification of viral DNA sequences in simian virus 40-transformed Chinese hamster embryo cells. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6144–6148. doi: 10.1073/pnas.78.10.6144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Mariani B. D., Schimke R. T. Gene amplification in a single cell cycle in Chinese hamster ovary cells. J Biol Chem. 1984 Feb 10;259(3):1901–1910. [PubMed] [Google Scholar]
  11. PRITCHARD R. H., LARK K. G. INDUCTION OF REPLICATION BY THYMINE STARVATION AT THE CHROMOSOME ORIGIN IN ESCHERICHIA COLI. J Mol Biol. 1964 Aug;9:288–307. doi: 10.1016/s0022-2836(64)80208-4. [DOI] [PubMed] [Google Scholar]
  12. Rice G. C., Gray J. W., Dewey W. C. FACS analysis of a hyperthermia-induced alteration in Hoechst 33342 permeability and direct measurement of its relationship to cell survival. J Cell Physiol. 1985 Mar;122(3):387–396. doi: 10.1002/jcp.1041220308. [DOI] [PubMed] [Google Scholar]
  13. Rice G. C., Spiro I. J., Ling C. C. Detection of S-phase overreplication following chronic hypoxia using a monoclonal anti-BrdUrd. Int J Radiat Oncol Biol Phys. 1985 Oct;11(10):1817–1822. doi: 10.1016/0360-3016(85)90038-0. [DOI] [PubMed] [Google Scholar]
  14. Salmon S. E., Hamburger A. W., Soehnlen B., Durie B. G., Alberts D. S., Moon T. E. Quantitation of differential sensitivity of human-tumor stem cells to anticancer drugs. N Engl J Med. 1978 Jun 15;298(24):1321–1327. doi: 10.1056/NEJM197806152982401. [DOI] [PubMed] [Google Scholar]
  15. Schimke R. T. Gene amplification in cultured animal cells. Cell. 1984 Jul;37(3):705–713. doi: 10.1016/0092-8674(84)90406-9. [DOI] [PubMed] [Google Scholar]
  16. Schimke R. T., Sherwood S. W., Hill A. B., Johnston R. N. Overreplication and recombination of DNA in higher eukaryotes: potential consequences and biological implications. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2157–2161. doi: 10.1073/pnas.83.7.2157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Stark G. R., Wahl G. M. Gene amplification. Annu Rev Biochem. 1984;53:447–491. doi: 10.1146/annurev.bi.53.070184.002311. [DOI] [PubMed] [Google Scholar]
  18. THOMLINSON R. H., GRAY L. H. The histological structure of some human lung cancers and the possible implications for radiotherapy. Br J Cancer. 1955 Dec;9(4):539–549. doi: 10.1038/bjc.1955.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tannock I. F. The relation between cell proliferation and the vascular system in a transplanted mouse mammary tumour. Br J Cancer. 1968 Jun;22(2):258–273. doi: 10.1038/bjc.1968.34. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Thompson L. H., Fong S., Brookman K. Validation of conditions for efficient detection of HPRT and APRT mutations in suspension-cultured Chinese hamster ovary cells. Mutat Res. 1980 Feb;74(1):21–36. doi: 10.1016/0165-1161(80)90188-0. [DOI] [PubMed] [Google Scholar]
  21. Tlsty T. D., Brown P. C., Schimke R. T. UV radiation facilitates methotrexate resistance and amplification of the dihydrofolate reductase gene in cultured 3T6 mouse cells. Mol Cell Biol. 1984 Jun;4(6):1050–1056. doi: 10.1128/mcb.4.6.1050. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Varshavsky A. On the possibility of metabolic control of replicon "misfiring": relationship to emergence of malignant phenotypes in mammalian cell lineages. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3673–3677. doi: 10.1073/pnas.78.6.3673. [DOI] [PMC free article] [PubMed] [Google Scholar]

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