Enhanced Detection of Deleterious and Other Germline Mutations of hMSH2 and hMLH1 in Japanese Hereditary Nonpolyposis Colorectal Cancer Kindreds

Autor: Kokichi Sugano, Noriko Fukayama, Hidefumi Kashiwabara, Sachio Nomura, Shin Fujita, Takao Sekiya, Takahiro Taniguchi, Tadao Kakizoe, Seiji Ohhigashi, Takayuki Akasu, Yoshihiro Moriya
Rok vydání: 2000
Předmět:
congenital
hereditary
and neonatal diseases and abnormalities
Time Factors
Colorectal cancer
DNA Mutational Analysis
Nonsense-mediated decay
Biophysics
Biology
medicine.disease_cause
Biochemistry
chemistry.chemical_compound
Germline mutation
Gene Frequency
Japan
Proto-Oncogene Proteins
medicine
Humans
Coding region
Cycloheximide
Allele
Frameshift Mutation
Molecular Biology
Alleles
Germ-Line Mutation
Adaptor Proteins
Signal Transducing
Nucleic Acid Synthesis Inhibitors
Family Health
Protein Synthesis Inhibitors
Genetics
Mutation
Reverse Transcriptase Polymerase Chain Reaction
Nuclear Proteins
nutritional and metabolic diseases
Exons
Cell Biology
medicine.disease
Colorectal Neoplasms
Hereditary Nonpolyposis
digestive system diseases
Neoplasm Proteins
DNA-Binding Proteins
MutS Homolog 2 Protein
chemistry
Puromycin
RNA extraction
Carrier Proteins
MutL Protein Homolog 1
Anisomycin
Gene Deletion
Zdroj: Biochemical and Biophysical Research Communications. 271:120-129
ISSN: 0006-291X
Popis: Hereditary nonpolyposis colorectal cancer (HNPCC) is an autosomal, dominantly inherited cancer-prone syndrome. Here, we describe a novel and efficient approach for screening mutations of two major HNPCC susceptibility genes, hMSH2 and hMLH1. The system consists of RNA extraction from whole blood treated with the translation inhibitor, followed by long RT-PCR of the entire coding regions combined with direct sequencing. In analysis of 15 kindreds suspicious for HNPCC, 8 samples were subjected to analysis after puromycin treatment and 7 samples were analyzed without puromycin treatment. Three deleterious mutations were detected in the kindreds with puromycin treatment, while none were observed in those without puromycin. Signals from mutated alleles were enhanced after puromycin treatment and easily distinguished from the wild-type allele, achieved by suppression of nonsense-mediated mRNA decay. Furthermore, 12 other mutations were detected in 15 kindreds. The system is considered to be a reliable and useful approach for detecting germline mutations of hMSH2 and hMLH1 in HNPCC kindreds.
Databáze: OpenAIRE
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