title: Colon Cancer Cells Gene Expression Signature As Response to 5- Fluorouracil, Oxaliplatin, and Folinic Acid Treatment.
abstract: 5-FU cytotoxicity mechanism has been assigned both to the miss-incorporation of fluoronucleotides into RNA and DNA and to the inhibition of thymidylate synthase. 5-FU is one of the most widely used chemotherapeutic drugs, although it has severe side effects that may vary between patients. Pharmacogenetic studies related to 5-FU have been traditionally focused on the rate-limiting catabolic enzyme, dihydropyrimidine dehydrogenase that breaks 80-85% of 5-FU into its inactive metabolite. Choosing the right dosing scheme and chemotherapy strategy for each individual patient remains challenging for personalized chemotherapy management. In the general effort toward reduction of colorectal cancer mortality, in vitro screening studies play a very important role. To accelerate translation research, increasing interest has been focused on using in vivo-like models such as three-dimensional spheroids. The development of higher throughput assays to quantify phenotypic changes in spheroids is an active research area. Consequently, in this study we used the microarray technology to reveal the HT-29 colorectal adenocarcinoma cells gene expression signature as response to 5-FU/OXP/FA treatment in a state of the art 3D culture system. We report here an increased reactive oxygen species production under treatment, correlated with a decrease in cell viability and proliferation potential. With respect to the HT-29 cells gene expression under the treatment with 5-FU/OXP/FA, we found 15.247 genes that were significantly differentially expressed (p < 0.05) with a fold change higher that two-fold. Among these, 7136 genes were upregulated and 8111 genes were downregulated under experimental conditions as compared to untreated cells. The most relevant and statistic significant (p < 0.01) pathways in the experiment are associated with the genes that displayed significant differential expression and are related to intracellular signaling, oxidative stress, apoptosis, and cancer.
authors.affiliation: Department of Toxicology, Faculty of Pharmacy, "Carol Davila" University of Medicine and Pharmacy Bucharest, Romania.
authors.forename: Carolina
authors.initials: C
authors.lastname: Negrei
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Bucharest Bucharest, Romania.
authors.forename: Ariana
authors.initials: A
authors.lastname: Hudita
authors.affiliation: Department of Surgery, "Sf. Ioan" Clinical Emergency HospitalBucharest, Romania; Department II, Faculty of Dental Medicine, "Carol Davila" University of Medicine and PharmacyBucharest, Romania.
authors.forename: Octav
authors.initials: O
authors.lastname: Ginghina
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Bucharest Bucharest, Romania.
authors.forename: Bianca
authors.initials: B
authors.lastname: Galateanu
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Bucharest Bucharest, Romania.
authors.forename: Sorina Nicoleta
authors.initials: SN
authors.lastname: Voicu
authors.affiliation: Department of Toxicology, Faculty of Pharmacy, "Carol Davila" University of Medicine and Pharmacy Bucharest, Romania.
authors.forename: Miriana
authors.initials: M
authors.lastname: Stan
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Bucharest Bucharest, Romania.
authors.forename: Marieta
authors.initials: M
authors.lastname: Costache
authors.affiliation: Occupational Medicine Section, Department of Biomedical and Dental Sciences and Morphofunctional Imaging, University of Messina Messina, Italy.
authors.forename: Concettina
authors.initials: C
authors.lastname: Fenga
authors.affiliation: Research Laboratory of Clinical Pharmacology and Pharmacogenomics, School of Health Sciences, Faculty of Pharmacy, National and Kapodistrian University of Athens Athens, Greece.
authors.forename: Nikolaos
authors.initials: N
authors.lastname: Drakoulis
authors.affiliation: Department of Toxicology and Forensic Sciences, Medical School, University of Crete Heraklion, Greece.
authors.forename: Aristidis M
authors.initials: AM
authors.lastname: Tsatsakis
keywords: 5-fluorouracil; DPD polymorphism; colorectal cancer; dihydropyrimidine dehydrogenase; gene expression; oxaliplatin; personalized medicine