DNA methylation analysis using bisulfite treatment and PCR-single-strand conformation polymorphism in colorectal cancer showing microsatellite instability

Citation
M. Maekawa et al., DNA methylation analysis using bisulfite treatment and PCR-single-strand conformation polymorphism in colorectal cancer showing microsatellite instability, BIOC BIOP R, 262(3), 1999, pp. 671-676
Citations number
28
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biochemistry & Biophysics
Journal title
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
ISSN journal
0006-291X → ACNP
Volume
262
Issue
3
Year of publication
1999
Pages
671 - 676
Database
ISI
SICI code
0006-291X(19990907)262:3<671:DMAUBT>2.0.ZU;2-J
Abstract
The combination of bisulfite treatment and PCR-single-strand DNA conformati on polymorphism (SSCP) analysis is proposed for quantitative methylation as say, We applied this procedure to the methylation analysis of the hMLH1 pro moter region in colorectal cancer. An analysis of mixtures of known amounts of methylated and unmethylated DNA revealed a linear relation. Using a cal ibration curve, proportions of methylated DNA were calculated, The hMLH1 pr omoter region was highly methylated in about 80% of microsatellite instabil ity (MSI) (+) colorectal cancers, but in none of the MSI(-) colorectal canc ers. A significant correlation existed between hypermethylation of the hMLH 1 promoter and MSI, as in previous reports. In conclusion, bisulfite-PCR-SS CP (BiPS) analysis could be applied to the rapid identification of methylat ion status in multiple samples, quantification of methylation differences, and detection of methylation heterogeneity in amplified DNA fragments. (C) 1999 Academic Press.