MUTATIONS IN THE P53 GENE IN HUMAN ASTROCYTOMAS - DETECTION BY SINGLE-STRAND CONFORMATION POLYMORPHISM ANALYSIS AND DIRECT DNA-SEQUENCING

Citation
Sb. Hunter et al., MUTATIONS IN THE P53 GENE IN HUMAN ASTROCYTOMAS - DETECTION BY SINGLE-STRAND CONFORMATION POLYMORPHISM ANALYSIS AND DIRECT DNA-SEQUENCING, Modern pathology, 6(4), 1993, pp. 442-445
Citations number
26
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Pathology
Journal title
ISSN journal
0893-3952
Volume
6
Issue
4
Year of publication
1993
Pages
442 - 445
Database
ISI
SICI code
0893-3952(1993)6:4<442:MITPGI>2.0.ZU;2-1
Abstract
The p53 gene was examined in a series of formalin-fixed paraffin-embed ded astrocytic neoplasms of various types by polymerase chain reaction (PCR), single-strand conformation polymorphism analysis (SSCP), and d irect sequencing of amplified DNA. PCR primers were designed to amplif y three DNA fragments encompassing exons 5, 7, and 8 with splice sites , including all four mutational ''hot spots'' within this gene. SSCP w as performed in a polyacrylamide gel containing 10% glycerol. Two muta tions were found among the 20 high and intermediate grade adult astroc ytomas studied by this sensitive screening technique and confirmed by sequencing of the PCR product. (1) An anaplastic astrocytoma disclosed a T-A transversion in Codon 246 giving rise to a methionine to lysine amino acid substitution. (2) A giant cell glioblastoma disclosed a G to A transition in Codon 285 resulting in a glutamic acid to lysine su bstitution. Both mutations were associated with loss of the normal all ele. Twenty-three DNA fragments that disclosed no mutation by SSCP ana lysis were confirmed to be negative by direct sequencing of amplified DNA. No mutations were detected in a series of eight juvenile cerebell ar astrocytomas, a biologically distinct form of low-grade astrocytoma . Mutations of the p53 gene may play an important pathogenetic role in a subset of human astrocytomas.