PMID: 7547246Oct 1, 1995Paper

A microdissection approach to detect molecular markers during progression of prostate cancer

British Journal of Cancer
P BerthonN J Maitland

Abstract

To investigate the underlying mechanisms of carcinogenesis, we have developed a technique to determine the frequency of genetic changes in prostatic carcinoma tissue. We have demonstrated that at a ratio of between 1:4 and 1:9 mutant-normal alleles, the signal from a mutant TP53 allele is not apparent after polymerase chain reaction (PCR) amplification and further direct sequencing or single-strand conformation polymorphism (SSCP) analysis. To bypass this problem, which is inherent in the heterogeneity of the prostate tissue and of the tumour, we selected areas of graded prostate tumours (Gleason score) from cryosectioned preparations and microdissected these cells (20-100 cells). After anionic resin removal of proteins, PCR amplification of TP53 gene exons 5/6 and SSCP analysis, an abnormal SSCP band shift was observed in suspected tumour cells, compared with microdissected stromal cells used as an internal control, while (1) a crude preparation of tissue DNA carrying the tumour did not show any abnormality and (2) immunostaining by a set of monoclonal antibodies against TP53 protein remained negative. Nucleotide sequence analysis of the different bands confirmed the presence of a mutation in the TP53 gene exon 6 position 13,3...Continue Reading

Citations

Jun 13, 2000·Diagnostic Molecular Pathology : the American Journal of Surgical Pathology, Part B·J Thompson-HehirC Walker
Jul 11, 2006·Ultrastructural Pathology·Silvia Hernández, Josep Lloreta
Nov 9, 2005·Prostate Cancer and Prostatic Diseases·J K J DissD S Latchman
Aug 12, 2000·Redox Report : Communications in Free Radical Research·J K Burgess, R H Hazelton
Sep 2, 2010·PloS One·Regina Berretta, Pablo Moscato
Oct 1, 2014·Cancer Prevention Research·Tin Oo KhorAh-Ng Tony Kong
Jun 4, 2008·Cancer Research·Marta CamorianoDominic J Smiraglia

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