title: Molecular profile of 5-fluorouracil pathway genes in colorectal carcinoma.
abstract: This study addresses involvement of major 5-fluorouracil (5-FU) pathway genes in the prognosis of colorectal carcinoma patients.
  
  
  
  Testing set and two validation sets comprising paired tumor and adjacent mucosa tissue samples from 151 patients were used for transcript profiling of 15 5-FU pathway genes by quantitative real-time PCR and DNA methylation profiling by high resolution melting analysis. Intratumoral molecular profiles were correlated with clinical data of patients. Protein levels of two most relevant candidate markers were assessed by immunoblotting.
  
  
  
  Downregulation of DPYD and upregulation of PPAT, UMPS, RRM2, and SLC29A1 transcripts were found in tumors compared to adjacent mucosa in testing and validation sets of patients. Low RRM2 transcript level significantly associated with poor response to the first-line palliative 5-FU-based chemotherapy in the testing set and with poor disease-free interval of patients in the validation set irrespective of 5-FU treatment. UPP2 was strongly methylated while its transcript absent in both tumors and adjacent mucosa. DPYS methylation level was significantly higher in tumor tissues compared to adjacent mucosa samples. Low intratumoral level of UPB1 methylation was prognostic for poor disease-free interval of the patients (P = 0.0002). The rest of the studied 5-FU genes were not methylated in tumors or adjacent mucosa.
  
  
  
  The observed overexpression of several 5-FU activating genes and DPYD downregulation deduce that chemotherapy naïve colorectal tumors share favorable gene expression profile for 5-FU therapy. Low RRM2 transcript and UPB1 methylation levels present separate poor prognosis factors for colorectal carcinoma patients and should be further investigated.
authors.affiliation: Department of Toxicogenomics, National Institute of Public Health, Prague, Czech Republic.|Third Faculty of Medicine, Charles University, Prague, Czech Republic.|Biomedical Centre, Medical School Pilsen, Charles University in Prague, Pilsen, Czech Republic.
authors.forename: T
authors.initials: T
authors.lastname: Kunicka
authors.affiliation: Department of Molecular Biology of Cancer, Institute of Experimental Medicine, Czech Academy of Sciences, Videnska 1083, 142 00, Prague 4, Czech Republic.
authors.forename: P
authors.initials: P
authors.lastname: Prochazka
authors.affiliation: Department of Toxicogenomics, National Institute of Public Health, Prague, Czech Republic.
authors.forename: I
authors.initials: I
authors.lastname: Krus
authors.affiliation: Biomedical Centre, Medical School Pilsen, Charles University in Prague, Pilsen, Czech Republic.|Department of Molecular Biology of Cancer, Institute of Experimental Medicine, Czech Academy of Sciences, Videnska 1083, 142 00, Prague 4, Czech Republic.
authors.forename: P
authors.initials: P
authors.lastname: Bendova
authors.affiliation: Department of Toxicogenomics, National Institute of Public Health, Prague, Czech Republic.
authors.forename: M
authors.initials: M
authors.lastname: Protivova
authors.affiliation: Department of Toxicogenomics, National Institute of Public Health, Prague, Czech Republic.|Biomedical Centre, Medical School Pilsen, Charles University in Prague, Pilsen, Czech Republic.
authors.forename: S
authors.initials: S
authors.lastname: Susova
authors.affiliation: Department of Toxicogenomics, National Institute of Public Health, Prague, Czech Republic.|Third Faculty of Medicine, Charles University, Prague, Czech Republic.|Biomedical Centre, Medical School Pilsen, Charles University in Prague, Pilsen, Czech Republic.
authors.forename: V
authors.initials: V
authors.lastname: Hlavac
authors.affiliation: Biomedical Centre, Medical School Pilsen, Charles University in Prague, Pilsen, Czech Republic.|Deparment of Surgery, Teaching Hospital and Medical School Pilsen, Charles University in Prague, Pilsen, Czech Republic.
authors.forename: V
authors.initials: V
authors.lastname: Liska
authors.affiliation: Deparment of Surgery, Teaching Hospital and Medical School Pilsen, Charles University in Prague, Pilsen, Czech Republic.
authors.forename: P
authors.initials: P
authors.lastname: Novak
authors.affiliation: Department of Surgery, General University Hospital in Prague, First Medical Faculty, Charles University, Prague, Czech Republic.
authors.forename: M
authors.initials: M
authors.lastname: Schneiderova
authors.affiliation: Biomedical Centre, Medical School Pilsen, Charles University in Prague, Pilsen, Czech Republic.
authors.forename: P
authors.initials: P
authors.lastname: Pitule
authors.affiliation: Biomedical Centre, Medical School Pilsen, Charles University in Prague, Pilsen, Czech Republic.|Deparment of Surgery, Teaching Hospital and Medical School Pilsen, Charles University in Prague, Pilsen, Czech Republic.
authors.forename: J
authors.initials: J
authors.lastname: Bruha
authors.affiliation: Biomedical Centre, Medical School Pilsen, Charles University in Prague, Pilsen, Czech Republic.|Deparment of Surgery, Teaching Hospital and Medical School Pilsen, Charles University in Prague, Pilsen, Czech Republic.
authors.forename: O
authors.initials: O
authors.lastname: Vycital
authors.affiliation: Biomedical Centre, Medical School Pilsen, Charles University in Prague, Pilsen, Czech Republic. pvodicka@biomed.cas.cz.|Department of Molecular Biology of Cancer, Institute of Experimental Medicine, Czech Academy of Sciences, Videnska 1083, 142 00, Prague 4, Czech Republic. pvodicka@biomed.cas.cz.
authors.forename: P
authors.initials: P
authors.lastname: Vodicka
authors.affiliation: Department of Toxicogenomics, National Institute of Public Health, Prague, Czech Republic. pavel.soucek@szu.cz.|Biomedical Centre, Medical School Pilsen, Charles University in Prague, Pilsen, Czech Republic. pavel.soucek@szu.cz.|Toxicogenomics Unit, National Institute of Public Health, Srobarova 48, 100 42, Prague 10, Czech Republic. pavel.soucek@szu.cz.
authors.forename: P
authors.initials: P
authors.lastname: Soucek
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology*; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000401:mortality / Q000473:pathology; D018899:CpG Islands; D019175:DNA Methylation; D005260:Female; D005472:Fluorouracil / Q000494:pharmacology*; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D006801:Humans; D008297:Male; D008875:Middle Aged; D060787:Neoplasm Grading; D009367:Neoplasm Staging; D011401:Promoter Regions, Genetic; D012262:Ribonucleoside Diphosphate Reductase / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D059467:Transcriptome
chemical_list: D000964:Antimetabolites, Antineoplastic; C093713:ribonucleotide reductase M2; D012262:Ribonucleoside Diphosphate Reductase; D005472:Fluorouracil
keywords: 5-fluorouracil; Colorectal carcinoma; Expression; Methylation; Prognosis