title: DNA mismatch repair gene mutations in 55 kindreds with verified or putative hereditary non-polyposis colorectal cancer.
abstract: The DNA mismatch repair genes MSH2 and MLH1 have been shown to account for a major share of hereditary non-polyposis colorectal cancer (HNPCC). We searched for germline mutations in these genes in 35 HNPCC kindreds fulfilling the Amsterdam diagnostic criteria and in a further 20 kindreds with an average of four affected members per family but not meeting the formal criteria. We first screened for truncations by reverse transcriptase (RT)-PCR. If no mutation was found, we screened genomic DNA by a novel application of two-dimensional (2-D) DNA electrophoresis that allows the simultaneous study of all exons of each gene. All abnormalities were followed up by sequencing. Eight different pathogenic germline mutations were found, two in MSH2 and six in MLH1. We report three major conclusions. First, these mutations together accounted for 86% (30/35) of the kindreds meeting the Amsterdam criteria, but only 30% (6/20) of the remaining kindreds, suggesting differences in etiology. Second, MLH1 was involved in > 90% (34/36) of kindreds with a known predisposing mutation, suggesting that mutations in the MLH1 gene are responsible for most HNPCC kindreds in Finland. Third, our results indicate that the successive application of RT-PCR and 2-D DNA electrophoresis is a sensitive and efficient method for mutation screening in typical HNPCC.
authors.affiliation: Department of Medical Genetics, Haartman Institute, University of Helsinki, Finland.
authors.forename: M
authors.initials: M
authors.lastname: Nyström-Lahti
authors.forename: Y
authors.initials: Y
authors.lastname: Wu
authors.forename: A L
authors.initials: AL
authors.lastname: Moisio
authors.forename: R M
authors.initials: RM
authors.lastname: Hofstra
authors.forename: J
authors.initials: J
authors.lastname: Osinga
authors.forename: J P
authors.initials: JP
authors.lastname: Mecklin
authors.forename: H J
authors.initials: HJ
authors.lastname: Järvinen
authors.forename: J
authors.initials: J
authors.lastname: Leisti
authors.forename: C H
authors.initials: CH
authors.lastname: Buys
authors.forename: A
authors.initials: A
authors.lastname: de la Chapelle
authors.forename: P
authors.initials: P
authors.lastname: Peltomäki
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D000328:Adult; D003123:Colorectal Neoplasms, Hereditary Nonpolyposis / Q000235:genetics* / Q000378:metabolism; D004252:DNA Mutational Analysis; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000235:genetics*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005656:Fungal Proteins / Q000235:genetics*; D005820:Genetic Testing; D014644:Genetic Variation; D006801:Humans; D000070957:MutL Protein Homolog 1; D051718:MutS Homolog 2 Protein; D016133:Polymerase Chain Reaction / Q000379:methods*; D029701:Saccharomyces cerevisiae Proteins
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C088859:MLH1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D000070957:MutL Protein Homolog 1; D051718:MutS Homolog 2 Protein