title: Gemcitabine and cytosine arabinoside cytotoxicity: association with lymphoblastoid cell expression.
abstract: Two cytidine analogues, gemcitabine (dFdC) and 1-beta-d-arabinofuranosylcytosine (AraC), show significant therapeutic effect in a variety of cancers. However, response to these drugs varies widely. Evidence from tumor biopsy samples shows that expression levels for genes involved in the cytidine transport, metabolism, and bioactivation pathway contribute to this variation in response. In the present study, we set out to test the hypothesis that variation in gene expression both within and outside of this "pathway" might influence sensitivity to gemcitabine and AraC. Specifically, Affymetrix U133 Plus 2.0 GeneChip and cytotoxicity assays were performed to obtain basal mRNA expression and IC(50) values for both drugs in 197 ethnically defined Human Variation Panel lymphoblastoid cell lines. Genes with a high degree of association with IC(50) values were involved mainly in cell death, cancer, cell cycle, and nucleic acid metabolism pathways. We validated selected significant genes by performing real-time quantitative reverse transcription-PCR and selected two representative candidates, NT5C3 (within the pathway) and FKBP5 (outside of the pathway), for functional validation. Those studies showed that down-regulation of NT5C3 and FKBP5 altered tumor cell sensitivity to both drugs. Our results suggest that cell-based model system studies, when combined with complementary functional characterization, may help to identify biomarkers for response to chemotherapy with these cytidine analogues.
authors.affiliation: Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, Rochester, Minnesota 55905, USA.
authors.forename: Liang
authors.initials: L
authors.lastname: Li
authors.forename: Brooke
authors.initials: B
authors.lastname: Fridley
authors.forename: Krishna
authors.initials: K
authors.lastname: Kalari
authors.forename: Gregory
authors.initials: G
authors.lastname: Jenkins
authors.forename: Anthony
authors.initials: A
authors.lastname: Batzler
authors.forename: Stephanie
authors.initials: S
authors.lastname: Safgren
authors.forename: Michelle
authors.initials: M
authors.lastname: Hildebrandt
authors.forename: Matthew
authors.initials: M
authors.lastname: Ames
authors.forename: Daniel
authors.initials: D
authors.lastname: Schaid
authors.forename: Liewei
authors.initials: L
authors.lastname: Wang