Mutational Analysis of Gastrointestinal Stromal Tumors (GISTs): Procedural Approach for Diagnostic Purposes

Cancer Genomics - Proteomics

SALVATORE CONTI⁶  MARIA LAURA DE MARCHIS⁵  RENATO COVELLO³  MARIO ROSELLI¹  RAFFAELE PALMIROTTA⁵  GIORGIA LUDOVICI⁵  BARBARA LEONE⁵  DAVID DELLA-MORTE²  FIORELLA GUADAGNI⁵  LEOPOLDO COSTARELLI⁴  PATRIZIA FERRONI⁵ 

[1] Medical Oncology Unit, Tor Vergata Clinical Center, University of Rome, Rome, ItalyMedical Oncology Unit, Tor Vergata Clinical Center, University of Rome, Rome, ItalyMedical Oncology Unit, Tor Vergata Clinical Center, University of Rome, Rome, Italy;Department of Advanced Biotechnologies and Bioimaging, Institute of Care and Scientific Research (IRCCS) San Raffaele Pisana, Rome, ItalyDepartment of Neurology, Miller School of Medicine, University of Miami, Miami, FL, U.S.A.Department of Advanced Biotechnologies and Bioimaging, Institute of Care and Scientific Research (IRCCS) San Raffaele Pisana, Rome, ItalyDepartment of Advanced Biotechnologies and Bioimaging, Institute of Care and Scientific Research (IRCCS) San Raffaele Pisana, Rome, ItalyDepartment of Neurology, Miller School of Medicine, University of Miami, Miami, FL, U.S.A.Department of Neurology, Miller School of Medicine, University of Miami, Miami, FL, U.S.A.Department of Advanced Biotechnologies and Bioimaging, Institute of Care and Scientific Research (IRCCS) San Raffaele Pisana, Rome, ItalyDepartment of Neurology, Miller School of Medicine, University of Miami, Miami, FL, U.S.A.;Department of Pathology, “Regina Elena” National Cancer Institute Rome, ItalyDepartment of Pathology, “Regina Elena” National Cancer Institute Rome, ItalyDepartment of Pathology, “Regina Elena” National Cancer Institute Rome, Italy;Department of Pathology, San Giovanni Addolorata Hospital, Rome, Italy.Department of Pathology, San Giovanni Addolorata Hospital, Rome, Italy.Department of Pathology, San Giovanni Addolorata Hospital, Rome, Italy.;Department of Advanced Biotechnologies and Bioimaging, Institute of Care and Scientific Research (IRCCS) San Raffaele Pisana, Rome, ItalyDepartment of Advanced Biotechnologies and Bioimaging, Institute of Care and Scientific Research (IRCCS) San Raffaele Pisana, Rome, ItalyDepartment of Advanced Biotechnologies and Bioimaging, Institute of Care and Scientific Research (IRCCS) San Raffaele Pisana, Rome, Italy;Genechron Laboratory - C.R. ENEA Casaccia, Rome, ItalyGenechron Laboratory - C.R. ENEA Casaccia, Rome, ItalyGenechron Laboratory - C.R. ENEA Casaccia, Rome, Italy

关键词: Gastrointestinal stromal tumors;    KIT;    PDGFRA;    mutational analysis;    pharmacogenomics;   

DOI  :  

来源: Delinasios GJ CO

【 摘 要 】

Background: Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors in the digestive tract characterized, in the majority of cases, by activating mutations in the KIT (v-kit Hardy-Zuckerman 4 feline sarcoma viral oncogene homolog) or PDGFRA (platelet-derived growth factor receptor, alpha polypeptide) genes. Mutations affecting these tyrosine kinase receptors are also responsible for the mechanisms of primary and secondary drug resistance during the treatment with tyrosine kinase inhibitors. We performed mutational analysis to evaluate the pharmacotherapy susceptibility of GISTs, adopting a comprehensive procedural approach, in order to optimize the identification of mutations that may result in cellular resistance to conventional therapy. Materials and Methods: DNA from paraffin-embedded tumor sections from 40 GISTs were analyzed using microdissection, direct sequencing analysis and allelic separation by cloning. Results: KIT mutations were found in 55.0% of the tumor samples. PDGFRA mutations were present in 5.0% of cases. Allelic cloning assay allowed for better definition of the extent of the mutations and clarification of the exact nucleotidic position of complex mutations. Conclusion: Our experience suggests that sequential microdissection, direct sequencing and allelic separation by PCR cloning of large variants may improve the approach to mutational analysis and interpretation of sequence data of KIT and PDGFRA in patients with GIST.

【 授权许可】

Unknown