title: Detection of microsatellite instability in cancers by arbitrarily primed-PCR fingerprinting using a fluorescently labeled primer (FAP-PCR).
abstract: The microsatellite mutator phenotype (MMP), detected as a change in the number of repeating units in hundreds of thousands of microsatellite sequences in the tumor cell genome, underlies the carcinogenesis of a variety of tumors including sporadic and hereditary nonpolyposis colon cancers. This enhanced microsatellite instability was discovered using arbitrarily primed polymerase chain reaction (AP-PCR) fingerprinting of DNA from colon cancers. In this study, we found an arbitrary primer that can amplify multiple DNA fragments containing repeated sequences, including the poly A tracts found in the Alu repeats of the human genome. The combined use of primer labeling with fluorescence and an automated DNA sequencing analysis of AP-PCR products (FAP-PCR) detected alterations in fingerprint bands in all DNA samples previously determined to belong to the MMP. Fluorescent AP-PCR fingerprinting using this single arbitrary primer provides a convenient and efficient method for detecting tumor specific fingerprint alterations that are usually undetectable by conventional fingerprinting.
authors.affiliation: Oncogene Division, National Cancer Center Research Institute, Tokyo, Japan.
authors.forename: J
authors.initials: J
authors.lastname: Yasuda
authors.forename: H
authors.initials: H
authors.lastname: Kashiwabara
authors.forename: K
authors.initials: K
authors.lastname: Kawakami
authors.forename: K
authors.initials: K
authors.lastname: Uematsu
authors.forename: K
authors.initials: K
authors.lastname: Sugano
authors.forename: M
authors.initials: M
authors.lastname: Perucho
authors.forename: T
authors.initials: T
authors.lastname: Sekiya
mesh_terms: D015139:Blotting, Southern; D002877:Chromosomes, Human / Q000737:chemistry; D003001:Cloning, Molecular; D016172:DNA Fingerprinting; D017931:DNA Primers; D004273:DNA, Neoplasm / Q000032:analysis; D005456:Fluorescent Dyes; D006801:Humans; D018895:Microsatellite Repeats; D009369:Neoplasms / Q000378:metabolism*; D010641:Phenotype; D016133:Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D004273:DNA, Neoplasm; D005456:Fluorescent Dyes