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 |
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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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