A unique method for mutation analysis of tumor suppressor genes in colorectal carcinomas using a crypt isolation technique.

Abstract:

BACKGROUND:Contamination of nontumor tissue makes genetic analysis difficult. For this reason, it is important to obtain pure tumor tissue to ensure accurate genetic analysis. OBJECTIVE:To accurately assess the incidence of mutation of tumor suppressor genes (p53: exon 5-8; APC: mutated cluster region; NF-2 gene: all exons) in 45 colorectal carcinomas. METHODS:We developed an application of the polymerase chain reaction-single-strand conformation polymorphism and DNA sequence by coupling them with crypt isolation. RESULTS:Mutations of p53 and APC genes were found in 24 and 22 of 45 colorectal carcinomas, respectively. No mutation of the NF-2 gene was observed in this cancer. Single-strand conformation polymorphism using a crypt isolation technique showed a clear migrating band and no false-positive data. CONCLUSIONS:The crypt isolation technique is a useful method for accurately analyzing genetic alterations. Furthermore, our proposed method confirmed the morphological findings obtained before the genetic analysis.

journal_name

Arch Pathol Lab Med

authors

Sugai T,Habano W,Nakamura Si,Uesugi N,Sasou S,Itoh C

doi

10.1043/0003-9985(2000)124<0382:AUMFMA>2.0.CO;2

subject

Has Abstract

pub_date

2000-03-01 00:00:00

pages

382-6

issue

3

eissn

0003-9985

issn

1543-2165

journal_volume

124

pub_type

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