title: Frequency of p16(INK4A) alterations and K-ras mutations in intrahepatic cholangiocarcinoma of the liver.
abstract: BACKGROUND
  Inactivation of the tumour suppressor gene p16 (CDKN2/MTS-1/INK4A) and K-ras mutations are among the most frequent genetic alterations in human malignancies.
  
  
  OBJECTIVE
  To investigate the tumour suppressor gene p16 and its possible association with K-ras mutations in intrahepatic cholangiocarcinomas of the liver.
  
  
  METHODS
  The status of p16 was evaluated in 41 cholangiocarcinomas by methylation specific polymerase chain reaction, microsatellite analysis, DNA sequencing, and immunohistochemical staining. K-ras mutations were determined by direct DNA sequencing analyses after microdissection. The results obtained were correlated with histopathological variables and patient survival.
  
  
  RESULTS
  Hypermethylation of the 5' CpG island of the p16 gene was found in 34 of 41 (83%) carcinomas. Homozygous deletion at the p16 region was present in two (5%), and loss of heterozygosity (LOH) in eight cases (20%). We failed to detect p16 gene missense mutations. K-ras mutations were found in 22 of 41 (54%) cholangiocarcinomas and in two cases of tumour surrounding non-neoplastic liver tissue. All 22 cancers with K-ras mutations also exhibited methylated p16. We failed to observe a correlation between K-ras or p16 status and histopathological factors or prognosis of patients.
  
  
  CONCLUSIONS
  These data suggest that inactivation of the p16 gene is a frequent event in cholangiocarcinoma. The most common somatic alteration is promotor methylation of the p16 gene which is closely associated with K-ras mutations. We failed to establish p16 or K-ras status as independent prognostic factors in these tumours.
authors.affiliation: Institute of Pathology, University of Leipzig, Liebigstr 26, 04103 Leipzig, Germany. tana@medizin.uni-leipzig.de
authors.forename: A
authors.initials: A
authors.lastname: Tannapfel
authors.forename: M
authors.initials: M
authors.lastname: Benicke
authors.forename: A
authors.initials: A
authors.lastname: Katalinic
authors.forename: D
authors.initials: D
authors.lastname: Uhlmann
authors.forename: F
authors.initials: F
authors.lastname: Köckerling
authors.forename: J
authors.initials: J
authors.lastname: Hauss
authors.forename: C
authors.initials: C
authors.lastname: Wittekind
mesh_terms: D001650:Bile Duct Neoplasms / Q000235:genetics*; D001653:Bile Ducts, Intrahepatic; D018281:Cholangiocarcinoma / Q000235:genetics*; D018899:CpG Islands / Q000235:genetics; D019175:DNA Methylation; D017353:Gene Deletion; D019942:Genes, p16 / Q000235:genetics*; D011905:Genes, ras / Q000235:genetics*; D006801:Humans; D007150:Immunohistochemistry; D019656:Loss of Heterozygosity; D018895:Microsatellite Repeats; D009154:Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011379:Prognosis; D012189:Retrospective Studies; D017422:Sequence Analysis, DNA