title: Novel germline hMSH2 genomic deletion and somatic hMSH2 mutations in a hereditary nonpolyposis colorectal cancer family.
abstract: Germline and somatic mutations of the hMSH2 gene were determined in a Japanese hereditary nonpolyposis colorectal cancer (HNPCC) family fulfilling the Amsterdam criteria. PCR-SSCP-sequencing of genomic DNA detected a somatic hMSH2 mutation of an A deletion at codon 227-229 in a duodenal carcinoma and a somatic hMSH2 mutation of an A insertion at codon 21 in a gastric carcinoma from affected family members, both carcinomas exhibiting high microsatellite instability. However, no germline hMSH2 mutation was detected by the PCR-SSCP-sequencing method. Genomic DNA was then analyzed by Southern blot hybridization using three hMSH2 cDNA probes (probe A involving exons 1-5, probe B involving exons 4-11 and probe C involving exons 9-16) after digestion by restriction enzymes, EcoRI, HindIII and NsiI. The NsiI digest of DNA from normal tissues of affected members exhibited an aberrant 8.6 kb restriction fragment, in addition to the normal 10.6 kb fragment, when hybridized to probes A and B. This suggested the presence of a heterozygous 2kb genomic deletion encompassing exon 4, 5 or 6. RT-PCR-sequencing revealed that the deleted region encompassed exon 5. This novel genomic deletion of the hMSH2 gene was confirmed to be pathogenic, and the Southern hybridization pattern was applied to the pre-symptomatic diagnosis.
authors.affiliation: Hereditary Tumor Research Project, Tokyo Metropolitan Komagome Hospital, and Institute of Molecular Oncology, Showa University, Tokyo, Japan. mmiyaki@opal.famille.ne.jp
authors.forename: Michiko
authors.initials: M
authors.lastname: Miyaki
authors.forename: Takeru
authors.initials: T
authors.lastname: Iijima
authors.forename: Tatsuro
authors.initials: T
authors.lastname: Yamaguchi
authors.forename: Shuya
authors.initials: S
authors.lastname: Shirahama
authors.forename: Taichi
authors.initials: T
authors.lastname: Ito
authors.forename: Masamichi
authors.initials: M
authors.lastname: Yasuno
authors.forename: Takeo
authors.initials: T
authors.lastname: Mori
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D001483:Base Sequence; D015139:Blotting, Southern; D003123:Colorectal Neoplasms, Hereditary Nonpolyposis / Q000453:epidemiology / Q000235:genetics*; D004260:DNA Repair; D004273:DNA, Neoplasm / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics*; D004379:Duodenal Neoplasms / Q000453:epidemiology / Q000235:genetics*; D005260:Female; D017353:Gene Deletion; D018095:Germ-Line Mutation / Q000235:genetics*; D006801:Humans; D008297:Male; D018895:Microsatellite Repeats; D008875:Middle Aged; D008969:Molecular Sequence Data; D051718:MutS Homolog 2 Protein; D010375:Pedigree; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D012334:RNA, Neoplasm / Q000235:genetics; D015183:Restriction Mapping; D013274:Stomach Neoplasms / Q000453:epidemiology / Q000235:genetics*
chemical_list: D004273:DNA, Neoplasm; D004268:DNA-Binding Proteins; D011518:Proto-Oncogene Proteins; D012334:RNA, Neoplasm; C497172:MSH2 protein, human; D051718:MutS Homolog 2 Protein