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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1989 Dec;86(23):9441–9445. doi: 10.1073/pnas.86.23.9441

Differences in the rates of gene amplification in nontumorigenic and tumorigenic cell lines as measured by Luria-Delbrück fluctuation analysis.

T D Tlsty 1, B H Margolin 1, K Lum 1
PMCID: PMC298512  PMID: 2687881

Abstract

It has been hypothesized that genomic fluidity is an important component of tumorigenesis. Previous studies described the relationship between tumorigenicity and one marker for genomic fluidity, gene amplification. In this report, these studies are extended with the rat liver epithelial cell lines to show that: (i) the amplification in these cells arises in a spontaneous fashion in the population (i.e., the variants detected are not preexisting in the population), and (ii) the rate of spontaneous amplification (mutation), as measured by Luria-Delbrück fluctuation analysis, is significantly lower in the nontumorigenic cells than in the tumorigenic cells. The rate was estimated by using the Po method and the method of means. The rate of spontaneous amplification of the gene encoding the multifunctional protein CAD (containing the enzymatic activities carbamoyl-phosphate synthase, aspartate transcarbamylase, and dihydroorotase) in the highly tumorigenic cells was significantly greater than that for the nontumorigenic cells, reaching almost 1 x 10(-4) events per cell per generation. The rate of this mutagenic event is high compared to the rate of point mutations usually reported in mammalian cells, and its potential contribution to the tumorigenic process will be discussed.

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Selected References

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  1. 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]
  2. Capizzi R. L., Jameson J. W. A table for the estimation of the spontaneous mutation rate of cells in culture. Mutat Res. 1973 Jan;17(1):147–148. doi: 10.1016/0027-5107(73)90265-0. [DOI] [PubMed] [Google Scholar]
  3. Cifone M. A., Fidler I. J. Increasing metastatic potential is associated with increasing genetic instability of clones isolated from murine neoplasms. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6949–6952. doi: 10.1073/pnas.78.11.6949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cole J., Arlett C. F., Green M. H. The fluctuation test as a more sensitive system for determining induced mutation in L5178Y mouse lymphoma cells. Mutat Res. 1976 Dec;41(2-3):377–386. doi: 10.1016/0027-5107(76)90110-x. [DOI] [PubMed] [Google Scholar]
  5. Crawford B. D., Barrett J. C., Ts'o P. O. Neoplastic conversion of preneoplastic Syrian hamster cells: rate estimation by fluctuation analysis. Mol Cell Biol. 1983 May;3(5):931–945. doi: 10.1128/mcb.3.5.931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Elmore E., Kakunaga T., Barrett J. C. Comparison of spontaneous mutation rates of normal and chemically transformed human skin fibroblasts. Cancer Res. 1983 Apr;43(4):1650–1655. [PubMed] [Google Scholar]
  7. Goldberg S., Defendi V. Increased mutation rates in doubly viral transformed Chinese hamster cells. Somatic Cell Genet. 1979 Nov;5(6):887–895. doi: 10.1007/BF01542648. [DOI] [PubMed] [Google Scholar]
  8. Kaden D. A., Bardwell L., Newmark P., Anisowicz A., Skopek T. R., Sager R. High frequency of large spontaneous deletions of DNA in tumor-derived CHEF cells. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2306–2310. doi: 10.1073/pnas.86.7.2306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kempe T. D., Swyryd E. A., Bruist M., Stark G. R. Stable mutants of mammalian cells that overproduce the first three enzymes of pyrimidine nucleotide biosynthesis. Cell. 1976 Dec;9(4 Pt 1):541–550. doi: 10.1016/0092-8674(76)90036-2. [DOI] [PubMed] [Google Scholar]
  10. Kendal W. S., Frost P. Pitfalls and practice of Luria-Delbrück fluctuation analysis: a review. Cancer Res. 1988 Mar 1;48(5):1060–1065. [PubMed] [Google Scholar]
  11. Kendal W. S., Wang R. Y., Frost P. Spontaneous mutation rates in cloned murine tumors do not correlate with metastatic potential, whereas the prevalence of karyotypic abnormalities in the parental tumors does. Int J Cancer. 1987 Sep 15;40(3):408–413. doi: 10.1002/ijc.2910400321. [DOI] [PubMed] [Google Scholar]
  12. Li I. C., Fu J., Hung Y. T., Chu E. H. Estimation of mutation rates in cultured mammalian cells. Mutat Res. 1983 Oct;111(2):253–262. doi: 10.1016/0027-5107(83)90068-4. [DOI] [PubMed] [Google Scholar]
  13. Li I. C., Wu S. C., Fu J., Chu E. H. A deterministic approach for the estimation of mutation rates in cultured mammalian cells. Mutat Res. 1985 Mar;149(1):127–132. doi: 10.1016/0027-5107(85)90017-x. [DOI] [PubMed] [Google Scholar]
  14. Luria S. E., Delbrück M. Mutations of Bacteria from Virus Sensitivity to Virus Resistance. Genetics. 1943 Nov;28(6):491–511. doi: 10.1093/genetics/28.6.491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Newcombe H. B. Delayed Phenotypic Expression of Spontaneous Mutations in Escherichia Coli. Genetics. 1948 Sep;33(5):447–476. doi: 10.1093/genetics/33.5.447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nowell P. C. The clonal evolution of tumor cell populations. Science. 1976 Oct 1;194(4260):23–28. doi: 10.1126/science.959840. [DOI] [PubMed] [Google Scholar]
  17. Otto E., McCord S., Tlsty T. D. Increased incidence of CAD gene amplification in tumorigenic rat lines as an indicator of genomic instability of neoplastic cells. J Biol Chem. 1989 Feb 25;264(6):3390–3396. [PubMed] [Google Scholar]
  18. Sager R., Gadi I. K., Stephens L., Grabowy C. T. Gene amplification: an example of accelerated evolution in tumorigenic cells. Proc Natl Acad Sci U S A. 1985 Oct;82(20):7015–7019. doi: 10.1073/pnas.82.20.7015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sager R. Mutation rates and mutational spectra in tumorigenic cell lines. Cancer Surv. 1988;7(2):325–333. [PubMed] [Google Scholar]
  20. 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]
  21. Seeger R. C., Brodeur G. M., Sather H., Dalton A., Siegel S. E., Wong K. Y., Hammond D. Association of multiple copies of the N-myc oncogene with rapid progression of neuroblastomas. N Engl J Med. 1985 Oct 31;313(18):1111–1116. doi: 10.1056/NEJM198510313131802. [DOI] [PubMed] [Google Scholar]
  22. Seshadri R., Kutlaca R. J., Trainor K., Matthews C., Morley A. A. Mutation rate of normal and malignant human lymphocytes. Cancer Res. 1987 Jan 15;47(2):407–409. [PubMed] [Google Scholar]
  23. Slamon D. J., Clark G. M., Wong S. G., Levin W. J., Ullrich A., McGuire W. L. Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science. 1987 Jan 9;235(4785):177–182. doi: 10.1126/science.3798106. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Tsao M. S., Grisham J. W., Chou B. B., Smith J. D. Clonal isolation of populations of gamma-glutamyl transpeptidase-positive and -negative cells from rat liver epithelial cells chemically transformed in vitro. Cancer Res. 1985 Oct;45(10):5134–5138. [PubMed] [Google Scholar]
  26. Tsao M. S., Grisham J. W., Nelson K. G. Clonal analysis of tumorigenicity and paratumorigenic phenotypes in rat liver epithelial cells chemically transformed in vitro. Cancer Res. 1985 Oct;45(10):5139–5144. [PubMed] [Google Scholar]
  27. Wahl G. M., Padgett R. A., Stark G. R. Gene amplification causes overproduction of the first three enzymes of UMP synthesis in N-(phosphonacetyl)-L-aspartate-resistant hamster cells. J Biol Chem. 1979 Sep 10;254(17):8679–8689. [PubMed] [Google Scholar]
  28. Wong A. J., Ruppert J. M., Eggleston J., Hamilton S. R., Baylin S. B., Vogelstein B. Gene amplification of c-myc and N-myc in small cell carcinoma of the lung. Science. 1986 Jul 25;233(4762):461–464. doi: 10.1126/science.3014659. [DOI] [PubMed] [Google Scholar]

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