NDLI: Improving the detection of p53 mutations in breast cancer by use of the FASAY, a functional assay.

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Improving the detection of p53 mutations in breast cancer by use of the FASAY, a functional assay.

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Improving the detection of p53 mutations in breast cancer by use of the FASAY, a functional assay.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Duddy, P. M. Hanby, A. M. Barnes, D. M. Camplejohn, R. S.
Description	Country affiliation: United kingdom Author Affiliation: Duddy PM ( Richard Dimbleby Department of Cancer Research, Guy's King's and St Thomas' School of Medicine, St Thomas' Hospital, London, United Kingdom.)
Abstract	The aim of this investigation was to examine the ability of the yeast-based functional assay, the functional analysis for the separation of alleles in yeast (FASAY), to detect p53 mutations in breast cancers when compared with immunohistochemistry and automated sequencing of the whole p53 gene (exons 1-11). To achieve this, all three methods were carried out on a cohort of aggressive breast tumors. In those tumors, in which the FASAY analysis indicated the presence of a mutation, cDNA was extracted from red yeast colonies and was sequenced to identify the base change in the p53 gene. The FASAY detected all 24 mutations found in the series of 48 tumors, whereas initial automated sequencing of genomic DNA detected 18/24 mutations. A second round of automated sequencing carried out using an independent source of genomic DNA detected mutations in 3 of the 6 tumors that originally appeared to lack a mutation in genomic DNA. All but 1 of the mutations originally missed by sequencing of genomic DNA were point mutations. Five mutations in this series (21%) were outside the commonly investigated exons 5-8, reinforcing the need to extend sequencing beyond this region. Of 24 tumors, 14 had strong immunohistochemical staining, and all 14 had p53 mutations; the majority of mutations missed by immunohistochemistry produced a truncated protein. Strong staining was not seen in tumors lacking a p53 mutation. The FASAY proved to be a rapid, reliable, and effective method for identifying those breast tumors harboring p53 mutations.
File Format	HTM / HTML
ISSN	15251578
e-ISSN	19437811
Journal	The Journal of Molecular Diagnostics
Issue Number	3
Volume Number	2
Language	English
Publisher	Elsevier
Publisher Date	2000-08-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Molecular Biology Breast Neoplasms Genetics Genes, P53 Mutation Tumor Suppressor Protein P53 Metabolism Alleles Dna Mutational Analysis Dna, Complementary Exons Frameshift Mutation Histocytochemistry Point Mutation Polymorphism, Genetic Sequence Analysis, Dna Time Factors Transcriptional Activation Transformation, Genetic Yeasts Comparative Study Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Molecular Medicine Pathology and Forensic Medicine

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