Switch to
| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
1
|
| pubmed:dateCreated |
1996-12-13
|
| pubmed:abstractText |
AGG to AGT mutations in codon 249 of the p53 tumor-suppressor gene are frequently observed in hepatocellular carcinomas (HCC) from areas where exposure to aflatoxin B1 (AFB) occurs. We developed a sensitive allele-specific polymerase chain reaction (AS-PCR) assay to detect this point mutation in non-neoplastic human liver tissues. Three oligonucleotide primers, 1 specific for the mutant allele and 2 specific for the wild-type allele were used. The mutant allele primer differed from the wild-type allele due to a G-to-T transversion in its terminal 3' nucleotide. The first stage involved amplification of exon 7 of p53 followed by a selective amplification of mutant codon 249 sequences. This method allowed for the detection of a mutant codon 249 allele in the presence of as many as 105 copies of the wild-type allele and was 100-fold more sensitive than the restriction fragment length polymorphism-PCR technique. We have applied this AS-PCR protocol to examine codon 249 AGT transversion in tumor and matched non-tumor liver samples from North American patients with hepatitis and from Mozambiquan patients exposed to AFB. Mutations were detected in 5 of 6 samples of non-neoplastic liver from Mozambiquan patients, all of whom were HBsAg- or HBcAg-positive and AFB-exposed. In contrast, no mutations were detected in non-neoplastic liver from North American patients with either HBV- or HCV-derived hepatitis and cirrhosis. This procedure is a simple and powerful approach for screening p53 codon 249 AGT mutation in heterogeneous non-neoplastic hepatocyte populations.
|
| pubmed:language |
eng
|
| pubmed:journal | |
| pubmed:citationSubset |
IM
|
| pubmed:chemical | |
| pubmed:status |
MEDLINE
|
| pubmed:month |
Sep
|
| pubmed:issn |
0020-7136
|
| pubmed:author | |
| pubmed:issnType |
Print
|
| pubmed:day |
27
|
| pubmed:volume |
68
|
| pubmed:owner |
NLM
|
| pubmed:authorsComplete |
Y
|
| pubmed:pagination |
21-5
|
| pubmed:dateRevised |
2007-7-24
|
| pubmed:meshHeading |
pubmed-meshheading:8895534-Adult,
pubmed-meshheading:8895534-Aflatoxin B1,
pubmed-meshheading:8895534-Aged,
pubmed-meshheading:8895534-Alleles,
pubmed-meshheading:8895534-Base Sequence,
pubmed-meshheading:8895534-Codon,
pubmed-meshheading:8895534-Female,
pubmed-meshheading:8895534-Genes, p53,
pubmed-meshheading:8895534-Hepatitis B,
pubmed-meshheading:8895534-Humans,
pubmed-meshheading:8895534-Liver,
pubmed-meshheading:8895534-Liver Neoplasms,
pubmed-meshheading:8895534-Male,
pubmed-meshheading:8895534-Middle Aged,
pubmed-meshheading:8895534-Molecular Sequence Data,
pubmed-meshheading:8895534-Mozambique,
pubmed-meshheading:8895534-Point Mutation,
pubmed-meshheading:8895534-Polymerase Chain Reaction,
pubmed-meshheading:8895534-Polymorphism, Restriction Fragment Length
|
| pubmed:year |
1996
|
| pubmed:articleTitle |
Allele-specific PCR analysis of p53 codon 249 AGT transversion in liver tissues from patients with viral hepatitis.
|
| pubmed:affiliation |
Lady Davis Institute for Medical Research and Jewish General Hospital, Department of Medicine, McGill University, Montreal, Canada.
|
| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|