title: Frequency of uridine monophosphate synthase Gly(213)Ala polymorphism in Caucasian gastrointestinal cancer patients and healthy subjects, investigated by means of new, rapid genotyping assays.
abstract: OBJECTIVE
  Uridine monophosphate synthase (UMPS) is a fundamental enzyme in pyrimidine synthesis. A single-nucleotide polymorphism, a G-C transversion at the 638th nucleotide, was demonstrated to increase UMPS activity and suggested to have clinical effects. The aims of this study were to set up simple genotyping methods and investigate the UMPS 638G>C polymorphism in the Caucasian population.
  
  
  RESULTS
  Two hundred forty-one patients with gastrointestinal cancers and 189 healthy subjects were enrolled. Genomic DNA was extracted from peripheral blood. A polymerase chain reaction-restriction fragment length polymorphism (RFLP) method was implemented using a forward primer incorporating a mismatched base to produce an artificial restriction site and BsrI restriction enzyme digestion; a denaturing high performance liquid chromatography (DHPLC) method was developed to further speed up UMPS genotyping. A 153 bp UMPS gene fragment was successfully amplified and analyzed in all samples. RFLP and DHPLC results showed a 100% match and where confirmed by direct sequencing. UMPS genotype distribution was similar in patients with cancer and control subjects.
  
  
  CONCLUSIONS
  Although no association was detected between UMPS variants and gastrointestinal cancer risk in Caucasians, polymerase chain reaction-RFLP with BsrI digestion and DHPLC set up at 59°C are reliable and cost-effective methods to genotype UMPS.
authors.affiliation: Department of Oncology, Rovigo General Hospital, Rovigo, Italy. gusella.milena@azisanrovigo.it
authors.forename: Milena
authors.initials: M
authors.lastname: Gusella
authors.forename: Laura
authors.initials: L
authors.lastname: Bertolaso
authors.forename: Caterina
authors.initials: C
authors.lastname: Bolzonella
authors.forename: Felice
authors.initials: F
authors.lastname: Pasini
authors.forename: Roberto
authors.initials: R
authors.lastname: Padrini
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000409:Alanine / Q000235:genetics; D019943:Amino Acid Substitution / Q000235:genetics / Q000502:physiology; D002277:Carcinoma / Q000453:epidemiology / Q000208:ethnology / Q000235:genetics*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003362:Cost-Benefit Analysis; D004252:DNA Mutational Analysis / Q000191:economics / Q000379:methods; D005260:Female; D005770:Gastrointestinal Neoplasms / Q000453:epidemiology / Q000208:ethnology / Q000235:genetics*; D005787:Gene Frequency; D020022:Genetic Predisposition to Disease; D060005:Genotyping Techniques / Q000191:economics / Q000379:methods; D005998:Glycine / Q000235:genetics; D006262:Health; D006801:Humans; D008297:Male; D008875:Middle Aged; D009097:Multienzyme Complexes / Q000235:genetics*; D009962:Orotate Phosphoribosyltransferase / Q000235:genetics*; D009964:Orotidine-5'-Phosphate Decarboxylase / Q000235:genetics*; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide; D015203:Reproducibility of Results; D044465:White People / Q000235:genetics
chemical_list: D009097:Multienzyme Complexes; C027157:uridine 5'-monophosphate synthase; D009962:Orotate Phosphoribosyltransferase; D009964:Orotidine-5'-Phosphate Decarboxylase; D000409:Alanine; D005998:Glycine