Genetic Variation in Thymidylate Synthase Confers Resistance to 5-Fluorodeoxyuridine

  • C. Todd Hughey
  • Karen W. Barbour
  • Franklin G. Berger
  • Sondra H. Berger
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 339)

Abstract

The human colorectal tumor cell line HCT 116 was resistant to the cytotoxic effects of 5-fluorodeoxyuridine (FdUrd). The response to FdUrd was increased only slightly by the presence of 10 μM folinic acid (CF). HCT 116 formed FdUMP and CH2H4PteGlu polyglutamates after exposure to FdUrd and CF. The sensitivity to FdUrd correlated well with the extent of TS inhibition. The role of TS in the resistance of the cells to FdUrd was examined. HCT 116 expresses two TS enzymes, which differ in pI. The more basic TS has been detected in only HCT 116 cells. The other TS is identical in pI to the enzymes detected in other human cells. The variant TS differs from the common by His replacement of Tyr at residue 33. The variant TS exhibited a 3-fold lower affinity for FdUMP than the common TS. The enzymes co-expressed in HCT 116 exhibited an FdUMP binding constant similar to that of the variant TS. TS-deficient cells were transfected with cDNAs encoding the two TS polypeptides. Transfectants expressing the variant TS were more resistant to FdUrd cytotoxicity than cells expressing the common TS. Thus, the structural variation in TS reduced enzyme affinity for FdUMP and conferred resistance to FdUrd.

Keywords

Folinic Acid Separate Determination Ehrlich Ascites Carcinoma Cell Thymidylate Synthetase Human Colonic Tumor Cell Line 
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.

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Copyright information

© Springer Science+Business Media New York 1993

Authors and Affiliations

  • C. Todd Hughey
    • 1
  • Karen W. Barbour
    • 1
  • Franklin G. Berger
    • 2
  • Sondra H. Berger
    • 2
  1. 1.Biological SciencesUniversity of South CarolinaColumbiaUSA
  2. 2.Departments of Basic Pharmaceutical SciencesUniversity of South CarolinaColumbiaUSA

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