title: Screening of RET gene mutations in multiple endocrine neoplasia type-2 using conformation sensitive gel electrophoresis (CSGE).
abstract: Multiple endocrine neoplasia type 2 (MEN2) is an autosomal dominant inherited tumor syndrome caused by RET proto-oncogene germline mutations (RET). Here we tested the Conformation Sensitive Gel Electrophoresis (CSGE) as a screening method for RET hot-spot mutations. Seven MEN2 families were studied by direct sequencing analysis, CSGE and Single Strand Conformational Polymorphism (SSCP). Using CSGE/SSCP, we were able to detect four out of five types of RET mutations verified by sequencing analysis: Cys620Arg, Cys634Arg, Cys634Tyr, and Met918Thr, furthermore a missense substitution at codon 648 (Val648Ile). RET polymorphisms 691 and 769 were also verified. Data obtained using CSGE/SSCP were fully concordant. We conclude that CSGE showed to be a sensitive, fast, low-cost, and simple procedure to detect RET mutations in codons which are reported as the most prevalent RET variants (approximately 95%) in large MEN2 series. As to the Val804Met mutation, this method still needs to be optimized.
authors.affiliation: Endocrine Genetics Unit (LIM-25), Endocrinology, Department of Internal Medicine, Hospital das Clínicas, University of São Paulo School of Medicine, SP.
authors.forename: Marcelo A C G dos
authors.initials: MA
authors.lastname: Santos
authors.forename: Elisangela Pereira de S
authors.initials: EP
authors.lastname: Quedas
authors.forename: Rodrigo de Almeida
authors.initials: Rde A
authors.lastname: Toledo
authors.forename: Delmar M
authors.initials: DM
authors.lastname: Lourenço
authors.forename: Sergio Pereira de A
authors.initials: SP
authors.lastname: Toledo
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D005091:Exons; D005820:Genetic Testing; D018095:Germ-Line Mutation / Q000235:genetics; D006801:Humans; D018813:Multiple Endocrine Neoplasia Type 2a / Q000235:genetics*; D018807:Polymorphism, Single-Stranded Conformational; D000090063:Proto-Oncogene Mas; D051096:Proto-Oncogene Proteins c-ret / Q000235:genetics*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods
chemical_list: C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D051096:Proto-Oncogene Proteins c-ret