Abstract
We attempt to identify the plasma membrane transporter involved in the uptake of 5′-deoxy-5-fluorouridine (5′-DFUR), an intermediate metabolite of capecitabine. This novel oral fluoropyrimidine is used in cancer treatments and is a direct precursor of the cytostatic agent 5′-fluorouracil. We also examine the role of the transporter in 5′-DFUR cytotoxicity. The human concentrative nucleoside transporter (hCNT1) was cloned from human fetal liver and expressed in Xenopus laevis oocytes. The two-electrode voltage-clamp technique was used to demonstrate that 5′-DFUR, but not capecitabine or 5′-FU, is an hCNT1 substrate. Then, hCNT1 was heterologously expressed in the mammalian cell line Chinese hamster ovary-K1. Functional expression was demonstrated by monitoring transport of radiolabeled substrates and by using a monospecific polyclonal antibody generated against the transporter. hCNT1-expressing cells were more sensitive to 5′-DFUR than vector-transfected or wild-type cells. The sensitivity of the three cell types to other agents such as cisplatin or 5′-FU was identical. In conclusion, this study shows that 1) the pharmacological profile of a nucleoside transporter can be determined by an electrophysiological approach; 2) the hCNT1 transporter is involved in 5′-DFUR uptake; and 3) hCNT1 expression may increase cell sensitivity to 5′-DFUR treatment. This study also reports for the first time the generation of an antibody against hCNT1, which may be useful in the elucidation of the relationship between hCNT1 expression and tumor response to capecitabine treatment.
Footnotes
- Received October 11, 2000.
- Accepted January 31, 2001.
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Send reprint requests to: Dr. Marçal Pastor-Anglada Department of Biochemistry and Molecular Biology University of Barcelona Diagonal 645, E-08028 Barcelona, Spain. E-mail:mpastor{at}porthos.bio.ub.es
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This study was mainly funded by Grants SAF99–0115 and 2FD1997–1268 (Plan Nacional de I+D, Plan Nacional de Salud) (M.P.-A.) and partially supported by Programa Praxis XXI-F.C.T. (Portugal) (J.F.M) and PIUNA (Universidad de Navarra, Spain) (M.P.L.).
- The American Society for Pharmacology and Experimental Therapeutics
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