title: Mutation and methylation analysis of TP53 in adrenal carcinogenesis.
abstract: OBJECTIVE
  To investigate the role of coding region mutation and promoter hypermethylation of TP53 in adrenocortical cancer formation.
  
  
  METHODS
  Twenty sporadic adrenocortical cancers (ACCs) and five normal adrenal tissue samples were available for analysis. Coding region mutation of TP53 in 20 ACCs was examined by polymerase chain amplification using intronic primers for exons 2-11 and direct sequencing of the product. In 10 ACCs and five normal adrenal tissue specimens, methylation of the 16 CpG sites within the TP53 promoter was examined using bisulphite methylation sequencing.
  
  
  RESULTS
  Coding region mutation in TP53 was demonstrated in 5 of 20 ACCs. There were four mis-sense mutations and one frameshift mutation. Four of 5 patients with a TP53 mutation had metastases at diagnosis or detected soon thereafter and 3 of 4 died of disease within 12 months of surgical resection. No methylation was seen in the TP53 promoter in 10 ACC and the five normal adrenal tissues examined.
  
  
  CONCLUSIONS
  Coding region mutation in TP53 occurs in 25% of ACCs with a trend toward a poorer prognosis. Promoter methylation of TP53 is not present in ACC as a mechanism for tumour suppressor gene (TSG) inactivation and, therefore, other genes in the 17p13 region are implicated in adrenal carcinogenesis.
authors.affiliation: Cancer Genetics, Kolling Institute of Medical Research, Royal North Shore Hospital, St Leonards, NSW 2065, Australia. stansidhu@nebsc.com.au
authors.forename: S
authors.initials: S
authors.lastname: Sidhu
authors.forename: E
authors.initials: E
authors.lastname: Martin
authors.forename: C
authors.initials: C
authors.lastname: Gicquel
authors.forename: J
authors.initials: J
authors.lastname: Melki
authors.forename: S J
authors.initials: SJ
authors.lastname: Clark
authors.forename: P
authors.initials: P
authors.lastname: Campbell
authors.forename: C J
authors.initials: CJ
authors.lastname: Magarey
authors.forename: K M
authors.initials: KM
authors.lastname: Schulte
authors.forename: H D
authors.initials: HD
authors.lastname: Röher
authors.forename: L
authors.initials: L
authors.lastname: Delbridge
authors.forename: B G
authors.initials: BG
authors.lastname: Robinson
mesh_terms: D000310:Adrenal Gland Neoplasms / Q000235:genetics* / Q000378:metabolism*; D000328:Adult; D000368:Aged; D019175:DNA Methylation; D005260:Female; D016158:Genes, p53; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D016133:Polymerase Chain Reaction; D011379:Prognosis; D011401:Promoter Regions, Genetic; D017421:Sequence Analysis