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Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1990 Oct;81(10):1039–1044. doi: 10.1111/j.1349-7006.1990.tb03343.x

Kinetic Analysis of 5‐Fluorouracil Action against Various Cancer Cells

Makoto Inaba 1,✉, Junko Mitsuhashi 1, Shogo Ozawa 1,†
PMCID: PMC5917971  PMID: 2146239

Abstract

Based on our recent kinetic analysis, which made it possible to distinguish between the cell‐killing actions of cell cycle phase‐specific and non‐specific agents, we attempted to elucidate the actions of 5‐fluorouracil (FUra) on three different cancer cell lines. By colony‐forming assay, the concentrations of fluorouridine (FUrd), fluorodeoxyuridine (FdUrd) or FUra giving 90% cell kill (IC90) at various exposure times (texps) were obtained. With P388 cells, the curve of texps‐IC90 for FUrd on a log‐log scale was linear with a slope of — 1, which is typical for cell cycle phase‐nonspecific agents. In contrast, the curve for FdUrd showed a much steeper slope than —1, which is characteristic for cell cycle phase‐specific agents. We found that the curve for FUra was exactly the same as that for FUrd, indicating that the mode of FUra action on P388 leukemia is analogous to that of FUrd. Similar results were observed with human colon and renal cancer cell lines, HT‐29 and KU‐2, although when the cells were exposed to relatively low concentrations of FUra for a long time, a cell cycle phase‐specific action became evident.

Keywords: 5‐Fluorouracil, 5‐Fluorouracil nucleosides, Cell‐killing action, Kinetic analysis

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References

  • 1.Ozawa, S., Sugiyama, Y., Mitsuhashi, Y., Kobayashi, T. and Inaba, M.Cell killing action of cell cycle phase‐nonspecific antitumor agents is dependent on concentration‐time product. Cancer Chemother. Pharmacol., 21, 185–190 (1988). [DOI] [PubMed] [Google Scholar]
  • 2.Ozawa, S., Sugiyama, Y., Mitsuhashi, J. and Inaba, M.Kinetic analysis of cell killing effect induced by cytosine arabinoside and cisplatin in relation to cell cycle phase specificity in human colon cancer and Chinese hamser cells. Cancer Res., 49, 3823–3828 (1989). [PubMed] [Google Scholar]
  • 3.Wilkinson, D. S. and Pitot, H. C.Inhibition of ribosomal ribonucleic acid maturation in Novikoff hepatoma cells by 5‐fluorouracil and 5‐fluorouridine. J. Biol. Chem., 248, 63–68 (1973). [PubMed] [Google Scholar]
  • 4.Cory, J. G., Breland, J. C. and Carter, G. L.Effect of 5‐fluorouracil on RNA metabolism in Novikoff hepatoma cells. Cancer Res., 39, 4905–4913 (1979). [PubMed] [Google Scholar]
  • 5.Evans, R. M., Laskin, J. D. and Hakala, M. T.Assessment of growth‐limiting events caused by 5‐fluorouracil in mouse cells and in human cells. Cancer Res., 40, 4113–4122 (1980). [PubMed] [Google Scholar]
  • 6.Drewinko, B., Yang, L. Y., Ho, D. H. W., Benvenuto, J., Loo, T. L. and Freireich, E. J.Treatment of cultured human colon carcinoma cells with fluorinated pyrimidines. Cancer, 45, 1144–1158 (1980). [DOI] [PubMed] [Google Scholar]
  • 7.Kufe, D. W. and Major, P. P.5‐Fluorouracil incorporation into human breast carcinoma RNA correlates with cytotoxicity. J. Biol. Chem., 256, 9802–9805 (1981). [PubMed] [Google Scholar]
  • 8.Sawyer, R. C., Stolfi, R. L., Nayak, R. and Martin, D. S.Mechanism of cytotoxicity in 5‐fluorouracil chemotherapy of two murine solid tumors In: “Necleosides and Cancer Treatment — Rational Approaches to Antimetabolite, Selectivity and Modulation,” ed. Tattersall M. H. N. and Fox R. M., pp. 309–338 (1981). Academic Press, Inc., New York. [Google Scholar]
  • 9.Glazer, R. I. and Lloyd, L. S.Association of cell lethality with incorporation of 5‐fluorouracil and 5‐fluorouridine into nuclear RNA in human colon carcinoma cells in culture. Mol. Pharmacol., 21, 468–473 (1982). [PubMed] [Google Scholar]
  • 10.Cohen, M. B. and Glazer, R. I.Cytotoxicity and the inhibition of ribosomal RNA processing in human colon carcinoma cells. Mol. Pharmacol., 27, 308–313 (1985). [PubMed] [Google Scholar]
  • 11.Herrick, D. and Kufe, D. W.Lethality associated with incorporation of 5‐fluorouracil into preribosomal RNA. Mol. Pharmacol., 26, 135–140 (1984). [PubMed] [Google Scholar]
  • 12.Washtien, W. L.Comparison of 5‐fluorouracil metabolism in two human gastrointestinal tumor cell lines. Cancer Res., 44, 909–914 (1984). [PubMed] [Google Scholar]
  • 13.Akazawa, S., Kumai, R., Yoshida, K., Ayusawa, D., Shimizu, K. and Seno, T.The cytotoxicity of 5‐fluorouracil is due to its incorporation into RNA not its inhibition of thymidylate synthase as evidenced by the use of a mouse cell mutant deficient in thymidylate synthase. Jpn. J. Cancer Res., 77, 620–624 (1986). [PubMed] [Google Scholar]
  • 14.Kanamaru, R., Kakuta, H., Sato, T., Ichioka, C. and Wakui, A.The inhibitory effects of 5‐fluorouracil on the metabolism of preribosomal and ribosomal RNA in L‐1210 cells in vitro. Cancer Chemother. Pharmacol., 17, 43–46 (1986). [DOI] [PubMed] [Google Scholar]
  • 15.Takimoto, C. H., Cadman, E. C. and Armstrong, R. D.Precursor‐dependent differences in the incorporation of fluorouracil in RNA. Mol. Pharmacol., 29, 637–642 (1986). [PubMed] [Google Scholar]
  • 16.Takimoto, C. H., Tan, Y. Y., Cadman, E. C. and Armstrong, R. D.Correlation between ribosomal RNA production and RNA‐directed fluoropyrimidine cytotoxicity. Biochem. Pharmacol., 36, 3243–3248 (1987). [DOI] [PubMed] [Google Scholar]
  • 17.Klubes, P., Connelly, K., Cerna, I. and Mandel, H. G.Effects of 5‐fluorouracil on 5‐fluorodeoxyuridine 5′‐monophosphate and 2‐deoxyuridine 5′‐monophosphate pools, and DNA synthesis in solid mouse L1210 and rat Walker 256 tumors. Cancer Res., 38, 2325–2331 (1978). [PubMed] [Google Scholar]
  • 18.Houghton, J. A. and Houghton, P. J.On the mechanism of cytotoxicity of fluorinated pyrimidines in four human colon adenocarcinoma xenografts maintained in immune‐deprived mice. Cancer, 45, 1159–1167 (1980). [DOI] [PubMed] [Google Scholar]
  • 19.Houghton, J. A., Maroda, S. J.Jr., Phillips, J. O. and Houghton, P. J.Biochemical determinants of responsiveness to 5‐fluorouracil and its derivatives in xenografts of human colorectal adenocarcinomas in mice. Cancer Res., 41, 144–149 (1981). [PubMed] [Google Scholar]
  • 20.Houghton, J. A., Weiss, K. D., Williams, L. G., Torrance, P. M. and Houghton, P. J.Relationship between 5‐fluoro‐2′‐deoxyuridylate, 2′‐deoxyuridylate, and thymidylate synthase activity subsequent to 5‐fluorouracil administration, in xenografts of human colon adenocarcinomas. Biochem. Pharmacol., 35, 1351–1358 (1986). [DOI] [PubMed] [Google Scholar]
  • 21.Houghton, J. A., Torrance, P. M., Radparvar, S., Williams, L. G. and Houghton, P. J.Binding of 5‐fluorodeoxy‐uridylate to thymidylate synthase in human colon adenocarcinoma xenografts. Eur. J. Cancer Clin. Oncol., 22, 505–510 (1986). [DOI] [PubMed] [Google Scholar]
  • 22.Evans, R. M., Laskin, J. D. and Hakala, M. T.Effects of excess folates and deoxyinosine on the activity and site of action of 5‐fluorouracil. Cancer Res., 41, 3288–3295 (1981). [PubMed] [Google Scholar]
  • 23.Yin, M‐B., Zakrzewski, S. F. and Hakala, M. T.Relationship of cellular folate cofactor pools to the activity of 5‐fluorouracil. Mol. Pharmacol., 23, 190–197 (1983). [PubMed] [Google Scholar]
  • 24.Berger, S. H. and Hakala, M. T.Relationship of dUMP and free FdUMP pools to inhibition of thymidylate synthase by 5‐fluorouracil. Mol. Pharmacol., 25, 303–309 (1984). [PubMed] [Google Scholar]
  • 25.Spears, C. P., Shahinian, A. H., Moran, R. G., Heidelberger, C. and Corbett, T. H.In vivo kinetics of thymidylate synthetase inhibition in 5‐fluorouracil‐sensitive and ‐resistant murine colon adenocarcinomas. Cancer Res., 42, 450–456 (1982). [PubMed] [Google Scholar]
  • 26.Spears, C. P., Gustavsson, B. G., Mitchell, M. S., Spicer, D., Berne, M., Bernstein, L. and Danenberg, P. V.Thymidylate synthetase inhibition in malignant tumors and normal liver of patients given intravenous 5 ‐ fluorouracil. Cancer Res., 44, 4144–4150 (1984). [PubMed] [Google Scholar]
  • 27.Keyomarsi, K. and Moran, R. G.Mechanism of the cytotoxic synergism of fluoropyrimidines and folinic acid in mouse leukemic cells. J. Biol. Chem., 263, 14402–14409 (1988). [PubMed] [Google Scholar]
  • 28.Byuyan, B. K., Scheidt, L. G. and Fraser, T. J.Cell cycle phase specificity of antitumor agents. Cancer Res., 32, 398–407 (1972). [PubMed] [Google Scholar]
  • 29.Ardalan, B., Cooney, D. A., Jayaram, H. N., Carrico, C. K., Glazer, R. I., Macdonald, J. and Schein, P. S.Mechanisms of sensitivity and resistance of murine tumors to 5‐fluorouracil. Cancer Res., 40, 1431–1437 (1980). [PubMed] [Google Scholar]

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