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Induction of Resistance to Fluorodeoxyuridine Cytotoxicity and DNA Damage in Human Tumor Cells by Expression of Escherichia coli Deoxyuridinetriphosphatase¹

Departments of Pharmacology [C. E. C., L. A. P., J. M.] and Radiation Oncology [E. H. R., M. A. D., T. S. L.], University of Michigan Medical School, Ann Arbor, Michigan, 48109-0504

Recent studies from our laboratory suggested that, in some human colorectal tumor cell lines, sensitivity to fluorodeoxyuridine may depend upon the extent of dUTP accumulation that occurs following drug treatment and that elevation of dUTPase activity might be the basis for some instances of resistance to fluoropyrimidines. To test this model, we expressed Escherichia coli dUTPase in an established human tumor cell line (HT29) and measured the effect of this manipulation on response to fluorodeoxyuridine. As predicted, HT29 derivatives containing dUTPase activity 4–5-fold higher than controls were protected from fluorode-oxyuridine-induced loss of clonogenicity and from formation of DNA double strand breaks. These data provide the first direct evidence that alteration in a component of the uracil misincorporation/misrepair pathway can confer resistance to fluoropyrimidines in human tumor cells.

¹ This work was supported by NIH Grants CA42761 and CA56663 (J. M.), a Clinical Investigator award (K08-CA1590; E. H. R.), CA53440 (T. S. L.), GCRC Grant M01-RR00042, a Lawrence Upjohn fellowship (C. E. C.), and a Predoctoral Fellowship from the University of Michigan Medical School Cancer Research Committee (C. E. C.).

² Current address: Johns Hopkins Oncology Center, Oncology 2-109, 600 North Wolfe Street, Baltimore, MD 21287-5001.

³ To whom requests for reprints should be addressed, at 4701 Upjohn Center for Clinical Pharmacology, University of Michigan Medical School, Ann Arbor, MI 48109-0504.

Received 12/29/93. Accepted 3/18/94.

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