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Enhancement of the Incorporation of 5-Fluorodeoxyuridylate into DNA of HL-60 Cells by Metabolic Modulations¹

Nagoya National Hospital, Department of Medicine, 4-1-1, Naka-ku, Nagoya, 460 [M. T., K. K.], and Department of Biochemistry, Institute for Developmental Research, Aichi Prefecture Colony, Kasugai, Aichi, 480-03 [S. Y.], Japan

The exposure of HL-60 human promyelocytic leukemia cells to 0.5 µM 5-fluoro-2'-[³H]deoxyuridine (FdUrd) for 16 hr resulted in the incorporation of 5.14 ± 0.31 (S.D.) x 10^-7 mol FdUrd into DNA per mol of DNA nucleotide, which corresponds to 0.146 ± 0.082 pmol FdUrd per 10⁷ cells. Pretreatment with 50 µM deoxythymidine for 24 hr led to a 2.7-fold increase in the incorporation of this analogue into newly synthesized DNA during the ensuing 16-hr exposure to 0.5 µM [³H]FdUrd. Pretreatment with 0.5 µM methotrexate for 3 hr also increased the [³H]FdUrd incorporation into newly synthesized DNA approximately 5-fold. The coexistence of deoxythymidine or methotrexate with [³H]FdUrd, however, led to decreased incorporation of FdUrd into DNA.

More than 50% of the radioactivity in DNA separated by Cs₂SO₄ equilibrium density gradient centrifugation was proven to be fluorodeoxyuridylate by means of its binding to Lactobacillus casei deoxythymidine monophosphate synthetase.

¹ This study was supported in part by a grant-in-aid for Cancer Research from the University of Health and Welfare of Japan.

² To whom requests for reprints should be addressed, at Nagoya National Hospital, Hematological Disease Center, Nagoya, Japan.

Received 11/ 8/82. Accepted 8/ 8/83.