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Cellular Sources of Thymidine Nucleotides: Studies for PET

Fred Hutchinson Cancer Research Center, and Departments of Medicine and Biostatistics, University of Washington, Seattle, Washington

Correspondence: For reprints contact: Anthony Shields, MD, PhD, Fred Hutchinson Cancer Research Center, 1124 Columbia St., Seatle, WA 98104.

ABSTRACT

The relative utilization of endogenously synthesized thymidine nucleotides and exogenously supplied thymidine analog was compared in a number of mammalian cell lines, tissues, and tumors. To measure the relative utilization, cells were incubated in tissue culture media containing the thymidine analog [³H]-5-bromo-2'-deoxyuridine (BUDR). After extraction of the DNA, the degree of substitution of the thymidine by BUDR was determined by density gradient centrifugation. All the cell lines and tissues tested utilized both exogenous BUDR and endogenous thymidine sources to a similar extent. The relative utilization of the exogenous pathway could be manipulated by varying the exogenous concentration of BUDR. Our results demonstrate that one can predict the relative utilization of these two pathways and can calculate the effective specific activity of the intracellular thymidine nucleotide pool. Such information is needed in interpreting ¹¹C-labeled thymidine uptake as measured by positron emission tomography.

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