title: Usefulness of p53 gene mutations in the supernatant of bile for diagnosis of biliary tract carcinoma: comparison with K- ras mutation.
abstract: BACKGROUND
  The sensitivity of bile cytology for the diagnosis of biliary tract carcinoma (BTCa) is still low. In addition, the incidence of detection of genetic mutations in the bile of BTCa is not satisfactory yet. To improve the molecular diagnosis of BTCa, we analyzed p53 and K- ras mutations in DNA extracted from not only the sediment but the supernatant of bile samples.
  
  
  METHODS
  Polymerase chain reaction-single-strand conformation polymorphism and direct sequencing were used for analyses of p53 mutations in exons 5 through 8. K- ras mutations at codon 12 were examined by mutant allele-specific amplification.
  
  
  RESULTS
  In bile supernatant from patients with BTCa, p53 and K- ras mutations were detected in 50.0% (15/30) and 56.7% (17/30) of cases, respectively. The incidence of p53and K- ras mutations in the sediment was 33.3% and 43.3%, respectively. When a combination assay with both genes was used, molecular abnormalities were detected in 80.0% of cases, including 3 in which p53 alone was positive. In addition, either p53 or K- rasmutations were detected in 12 of 15 (80.0%) cases of BTCa in which the cytologic diagnoses were negative. p53 mutations were detected in neither supernatant nor sediment in 20 patients with cholelithiasis, although the incidence of K- ras mutations in the sediment was 20%.
  
  
  CONCLUSIONS
  The incidence of p53 and K- ras mutations is higher in the supernatant than in the sediment, and simultaneous analyses of p53 and K- ras in the two bile fractions could enhance the genetic diagnosis of BTCa. Notably, the specificity of p53 mutations for cancer was very high in bile samples, and the sensitivity was also relatively high.
authors.affiliation: Department of Internal Medicine and Medical Oncology, Cancer Research Institute, Kanazawa University, 13-1 Takara-machi, Kanazawa 920-8641, Japan.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Yamaguchi
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Watanabe
authors.forename: Koushiro
authors.initials: K
authors.lastname: Ohtsubo
authors.forename: Tokio
authors.initials: T
authors.lastname: Wakabayashi
authors.forename: Norio
authors.initials: N
authors.lastname: Sawabu
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001646:Bile / Q000737:chemistry*; D001661:Biliary Tract Neoplasms / Q000175:diagnosis* / Q000235:genetics; D002277:Carcinoma / Q000175:diagnosis* / Q000235:genetics; D003581:Cytodiagnosis; D005260:Female; D011905:Genes, ras / Q000235:genetics*; D005819:Genetic Markers; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000032:analysis / Q000235:genetics*
chemical_list: D005819:Genetic Markers; D016159:Tumor Suppressor Protein p53