Logo image
Gemcitabine Cytotoxicity: Interaction of Efflux and Deamination
Journal article   Open access

Gemcitabine Cytotoxicity: Interaction of Efflux and Deamination

Dan Rudin, Liang Li, Nifang Niu, Krishna R. Kalari, Judith A. Gilbert, Matthew M. Ames and Liewei Wang
Journal of drug metabolism & toxicology, Vol.2(107), 1000107
02/02/2011
DOI: 10.4172/2157-7609.1000107
PMCID: PMC3144579
PMID: 21804948
url
https://doi.org/10.4172/2157-7609.1000107View
Published (Version of record) Open Access

Abstract

Gemcitabine is a cytidine analogue used in the treatment of various solid tumors. Little is known about how gemcitabine and its metabolites are transported out of cells. We set out to study the efflux of gemcitabine and the possible consequences of that process in cancer cells. We observed the efflux of gemcitabine and its deaminated metabolite, 2’,2’-difluorodeoxyuridine (dFdU) using high performance liquid chromatography and tandem mass spectrometry (LC-MS/MS) after gemcitabine treatment. Non-selective ABCC-transport inhibition with probenecid significantly increased intracellular dFdU concentrations, with a similar trend observed with verapamil, a non-selective ABCB1 and ABCG2 transport inhibitor. Neither probenecid nor verapamil altered intracellular gemcitabine levels after the inhibition of deamination with tetrahydrourudine, suggesting that efflux of dFdU, but not gemcitabine, was mediated by ABC transporters. MTS assays showed that probenecid increased sensitivity to gemcitabine. While dFdU displayed little cytotoxicity, intracellular dFdU accumulation inhibited cytidine deaminase, resulting in increased gemcitabine levels and enhanced cytotoxicity. Knockdown of ABCC3, ABCC5 or ABCC10 individually did not significantly increase gemcitabine sensitivity, suggesting the involvement of multiple transporters. In summary, ABCC-mediated efflux may contribute to gemcitabine resistance through increased dFdU efflux that allows for the continuation of gemcitabine deamination. Reversing efflux-mediated gemcitabine resistance may require broad-based efflux inhibition.

Details

Metrics

5 Record Views
Logo image