title: Codon 249 mutation in exon 7 of p53 gene in plasma DNA: maybe a new early diagnostic marker of hepatocellular carcinoma in Qidong risk area, China.
abstract: OBJECTIVE
  One of the characteristics of hepatocellular carcinoma (HCC) in Qidong area is the selective mutation resulting in a serine substitution at codon 249 of the p53 gene (1, 20), and it has been identified as a "hotspot" mutation in heptocellular carcinomas occurring in populations exposed to aflatoxin and with high prevalence of hepatitis B virus carriers (2,3,9, 10,16,24). We evaluated in this paper whether this "hotspot" mutation could be detected in cell-free DNA circulating in plasma of patients with hepatocellular carcinoma and cirrhosis in Qidong, China, and tried to illustrate the significance of the detection of this molecular biomarker.
  
  
  METHODS
  We collected blood samples from 25 hepatocellular carcinoma patients, 20 cirrhotic patients and 30 healthy controls in Qidong area. DNA was extracted and purified from 200 microl of plasma from each sample. The 249(Ser) p53 mutation was detected by restriction digestion analysis and direct sequencing of exon-7 PCR products.
  
  
  RESULTS
  We found in exon 7 of p53 gene G-T transversion at the third base of codon 249 resulting 249(Arg) - 249(Ser) mutation in 10/25 (40 %) hepatocellular carcinoma cases, 4/20 (20 %) cirrhotics, and 2/30 (7 %) healthy controls. The adjusted odds ratio for having the mutation was 22.1(95 % CI, 3.2-91.7) for HCC cases compared to controls.
  
  
  CONCLUSIONS
  These data show that the 249(Ser) p53 mutation in plasma is strongly associated with hepatocellular carcinoma in Qidong patients. We found this mutation was also detected, although it was at a much lower frequency, in plasma DNA of Qidong cirrhotics and healthy controls; We consider that these findings, together with the usual method of HCC diagnosis, will give more information in early diagnosis of HCC, and 249(Ser) p53 mutation should be developed to a new early diagnostic marker for HCC.
authors.affiliation: The State Key Laboratory of Genetic Engineering, Fudan University, 200433, Shanghai, China.
authors.forename: Xing-Hua
authors.initials: XH
authors.lastname: Huang
authors.forename: Lu-Hong
authors.initials: LH
authors.lastname: Sun
authors.forename: Dong-Dong
authors.initials: DD
authors.lastname: Lu
authors.forename: Yan
authors.initials: Y
authors.lastname: Sun
authors.forename: Li-Jie
authors.initials: LJ
authors.lastname: Ma
authors.forename: Xi-Ran
authors.initials: XR
authors.lastname: Zhang
authors.forename: Jian
authors.initials: J
authors.lastname: Huang
authors.forename: Long
authors.initials: L
authors.lastname: Yu
mesh_terms: D000328:Adult; D000368:Aged; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000453:epidemiology / Q000235:genetics*; D002681:China / Q000453:epidemiology; D003062:Codon; D004247:DNA / Q000097:blood* / Q000235:genetics / Q000302:isolation & purification; D005091:Exons; D005260:Female; D016158:Genes, p53; D006514:Hepatitis B Surface Antigens / Q000032:analysis; D006801:Humans; D008113:Liver Neoplasms / Q000453:epidemiology / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D020125:Mutation, Missense; D016133:Polymerase Chain Reaction; D018570:Risk Assessment; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*; D000509:alpha-Fetoproteins / Q000032:analysis
chemical_list: D003062:Codon; D006514:Hepatitis B Surface Antigens; D016159:Tumor Suppressor Protein p53; D000509:alpha-Fetoproteins; D004247:DNA