In Vitro and In Vivo Inhibition of Thymidylate Synthase of Human Colon Cancer by 5-Fluorouracil

  • Godefridus J. Peters
  • Emile J. Laurensse
  • Cees J. van Groeningen
  • Sybren Meijer
  • Herbert M. Pinedo
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 253A)


Thymidylate synthase (TS) is a key enzyme in de novo synthesis of TMP (Fig. 1). TS catalyzes the conversion of dUMP to TMP for which 5, 10-methylenetetrahydrofolate (CH2THF) acts as the methyl-donor; the Km for dUMP is about 1–5 μM (1–5). Inhibition of TS by FdUMP is one of the main mechanisms for the action of 5-fluorouracil (5FU). Without preincubation FdUMP acts as a potent competitive inhibitor of TS with a Ki of about 1 nM. The Km/Ki ratio is about 1000. The inhibition of TS by FdUMP is mediated by the formation of a tight-binding covalent ternary complex of FdUMP with TS and CH THF. Retention of the inhibition of TS is mainly related to the FdUMP/dUMP ratio (6), the FdUMP binding to TS and the stabilization of the ternary complex by CH2 THF or one of its polyglutama-tes (6, 7, 9). In vitro, resistance against 5FU or its analog 5-fluoro-2′-deoxyuridine (FUdR) has been related to altered kinetics of TS with respect to Km values for dUMP and FdUMP binding (2, 5, 8), disturbed folate pools (9), and the level of enzyme before treatment (6, 8). Gene amplification of TS has been demonstrated for FUdR-resistant sub-cell lines (10, 11). Recently evidence for gene amplication has also been obtained in a patient with colon cancer who developed resistance against 5FU (12), while in breast cancer patients binding of FdUMP and the effect of CH2THF decreased during development of resistance (13).


Pyrimidine Nucleoside Primary Colorectal Tumor Potent Competitive Inhibitor Tritium Release Assay FdUMP Binding 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Rode W, Kulikowski T, Kedzienska B, Jastreboff M, Shugar D (1984). Biochem Phannacol 33, 2699–2705.CrossRefGoogle Scholar
  2. 2.
    Bapat AR, Zarov C, Danenberg PV (1983). J Biol Chem 258, 4130–4136PubMedGoogle Scholar
  3. 3.
    Danenberg PV (1977). Biochim Biophys Acta 473, 73–92PubMedGoogle Scholar
  4. 4.
    Peters GJ, Laurensse E, Leyva A, Pinedo HM (1987). Eur J Cancer Clin Oncol 23, 1869–1881.PubMedCrossRefGoogle Scholar
  5. 5.
    Pinedo HM, Peters GJ (1988). J Clin Oncol, in pressGoogle Scholar
  6. 6.
    Berger SH, Hakala MT (1984). Mol Phannacol 25, 303–309Google Scholar
  7. 7.
    Houghton JA, Maroda SJ, Philips JO, Houghton PJ (1981). Cancer Res 41 144–149PubMedGoogle Scholar
  8. 8.
    Priest DG, Ledford SE, Doig MT (1980). Biochem Pharmacol 29, 1549–1553PubMedCrossRefGoogle Scholar
  9. 9.
    Yin MB, Zakrzewski SF, Hakala MT (1983). Mol Pharmacol 23, 190–197PubMedGoogle Scholar
  10. 10.
    Berger SH, Jenh CH, Johnson LF, Berger FG (1985). Mol Pharmacol 28, 461–467PubMedGoogle Scholar
  11. 11.
    Jenh CH, Geyer PK, Baskin F, Johnson LF (1985). Mol Pharmacol 28, 80–85PubMedGoogle Scholar
  12. 12.
    Clark JL, Berger SH, Mittelman A, Berger FG (1987). Cancer Treatm Rep 71, 261–265Google Scholar
  13. 13.
    Allegra CJ, Chabner BA, Scholar PW, Bagley C, Drake JC, Lippman ME (1987). NCI Monographs 5, 199–202PubMedGoogle Scholar
  14. 14.
    Spears CP, Shahinian AH, Moran RG, Heidelberger C, Corbett TH (1982). Cancer Res 42, 450–456PubMedGoogle Scholar
  15. 15.
    Spears CP, Gustavsson BG, Mitchell MS, Spicer D, Berne M, Bernstein L, Danenberg PV (1984). Cancer Res 44, 4144–4150PubMedGoogle Scholar
  16. 16.
    Peters GJ, Laurensse E, Leyva A, Lankelma J, Pinedo HM (1986). Cancer Res 46, 20–28PubMedCrossRefGoogle Scholar
  17. 17.
    Peters GJ, Laurensse E, Leyva A, Pinedo HM (1986). Clin Chim Acta 158, 193–198PubMedCrossRefGoogle Scholar
  18. 18.
    Moran RG, Spears CP, Heidelberger C (1979). Proc Natl Acad Sci USA 76, 1456–1460PubMedCrossRefGoogle Scholar
  19. 19.
    Femandes DJ, Cranford SK (1984). Anal Biochem 142, 378CrossRefGoogle Scholar
  20. 20.
    Houghton JA, Radparvar S, Torrance PM, Williams LG, Houghton PJ (1987). Biochem Pharmacol 36, 1285–1289PubMedCrossRefGoogle Scholar
  21. 21.
    Peters GJ. Laurensse E, Leyva A, Lankelma J, Pinedo HM (1985). Proc AACR 26, 12 (Abstract 48)Google Scholar
  22. 22.
    Petrelli N, Herrera L, Rustum Y, Burke P, Creavan P, Stulc J, Emrich LJ, Mittelman A (1988). J Clin Oncol 5, 1559–1564Google Scholar
  23. 23.
    Nadal JC, Van Groeningen CJ, Pinedo HM, Peters GJ (1988). Invest New Drugs, in pressGoogle Scholar
  24. 24.
    Hashimoto Y, Shiotani T, Weber G (1987) Anal Biochem 67, 340–346CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1989

Authors and Affiliations

  • Godefridus J. Peters
    • 1
  • Emile J. Laurensse
    • 1
  • Cees J. van Groeningen
    • 1
  • Sybren Meijer
    • 1
  • Herbert M. Pinedo
    • 1
  1. 1.Departments of Oncology and SurgeryFree University HospitalAmsterdamThe Netherlands

Personalised recommendations