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Molecular analysis of the APC gene in 105 Dutch kindreds with familial adenomatous polyposis: 67 germline mutations identified by DGGE, PTT, and southern analysis
Author(s) -
van der Luijt Rob. B.,
Khan P. Meera,
Vasen Hans F. A.,
Tops Carli M. J.,
van LeeuwenCornelisse Inge S. J.,
Wijnen Juul Th.,
van der Klift Heleen M.,
Plug Rob J.,
Griffioen Gerrit,
Fodde Riccardo
Publication year - 1997
Publication title -
human mutation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 162
eISSN - 1098-1004
pISSN - 1059-7794
DOI - 10.1002/(sici)1098-1004(1997)9:1<7::aid-humu2>3.0.co;2-8
Subject(s) - familial adenomatous polyposis , biology , adenomatous polyposis coli , genetics , exon , germline mutation , gene , mutation , germline , point mutation , genetic analysis , microbiology and biotechnology , cancer , colorectal cancer
Abstract Germline mutations of the adenomatous polyposis coli ( APC ) gene are responsible for familial adenomatous polyposis (FAP), an autosomal dominant predisposition to colorectal cancer. We screened the entire coding region of the APC gene for mutations in an unselected series of 105 Dutch FAP kindreds. For the analysis of exons 1–14, we employed the GC‐clamped denaturing gradient gel electrophoresis (DGGE), while the large exon 15 was examined using the protein truncation test. Using this approach, we identified 65 pathogenic mutations in the above 105 apparently unrelated FAP families. The mutations were predominantly either frameshifts (39/65) or single base substitutions (18/65), resulting in premature stop codons. Mutations that would predict abnormal RNA splicing were identified in seven cases. In one of the families, a nonconservative amino acid change was found to segregate with the disease. In spite of the large number of APC mutations reported to date, we identified 27 novel germline mutations in our patients, which reiterates the great heterogeneity of the mutation spectrum in FAP. In addition to the point mutations identified in our patients, structural rearrangements of APC were found in two pedigrees, by Southern blot analysis. The present study indicates that the combined use of DGGE, protein truncation test, and Southern blot analysis offers an efficient strategy for the presymptomatic diagnosis of FAP by direct mutation detection. We found that the combined use of the currently available molecular approaches still fails to identify the underlying genetic defect in a significant subset of the FAP families. The possible causes for this limitation are discussed. © 1997 Wiley‐Liss, Inc.