TY - JOUR
T1 - Multiplex mutation screening by mass spectrometry in gastrointestinal stromal tumours
AU - Kang, Guhyun
AU - Lee, Jeeyun
AU - Jang, Ki Taek
AU - Beadling, Carol
AU - Corless, Chistopher L.
AU - Heinrich, Micheal C.
AU - Park, Joon Oh
AU - Kang, Won Ki
AU - Park, Cheol Keun
AU - Kim, Kyoung Mee
N1 - Funding Information:
was supported by grants from National Research Foundation of Korea (S-2010-0503-000-1) and Samsung Biomedical Research Institute (SBRI C-A9-203-1).
PY - 2012/8
Y1 - 2012/8
N2 - Aims: Clinical decision making and optimal clinical trial design based on cancer genetic information will be increasingly informed by the mutational status of multiple genes. Methods: We performed mutation screening on 22 fresh frozen gastrointestinal stromal tumours (GISTs) using a multiplexed oncogene screening panel with a mass spectroscopy readout (MassARRAY). The panel can detect 390 known mutations across 30 genes, including several known to contribute to intracellular signalling in cancers (BRAF, PIK3CA, KRAS HRAS, NRAS, AKT1, CTNNB1, GNAQ, CDK4, MAP2K1 and MAP2K2). Results: Direct Sanger sequencing confirmed that 16 cases (73%) harboured KIT mutations, affecting exon 11, 13 and 17, and the remaining six were wild-type for both KIT and PDGFRA. The sensitivity of the multiplexed oncogene screening panel was 100% for identifying missense mutations in KIT. Only 17% of the deletion mutations were detected, because the panel was not designed for detecting these. A substitution in FBX4 exon 1 (S8R), representing a germline single-nucleotide polymorphism, was observed in a case with KIT exon 11 missense mutation. No other mutations were identified, including in the six wild-type GISTs. Conclusions: Our results indicate that mutations other than KIT or PDGFRA are rare in GISTs. Although multiplex mutation screening by mass spectrometry detected missense mutations accurately, it is not sufficient to screen mutations because deletion mutations are common in GISTs.
AB - Aims: Clinical decision making and optimal clinical trial design based on cancer genetic information will be increasingly informed by the mutational status of multiple genes. Methods: We performed mutation screening on 22 fresh frozen gastrointestinal stromal tumours (GISTs) using a multiplexed oncogene screening panel with a mass spectroscopy readout (MassARRAY). The panel can detect 390 known mutations across 30 genes, including several known to contribute to intracellular signalling in cancers (BRAF, PIK3CA, KRAS HRAS, NRAS, AKT1, CTNNB1, GNAQ, CDK4, MAP2K1 and MAP2K2). Results: Direct Sanger sequencing confirmed that 16 cases (73%) harboured KIT mutations, affecting exon 11, 13 and 17, and the remaining six were wild-type for both KIT and PDGFRA. The sensitivity of the multiplexed oncogene screening panel was 100% for identifying missense mutations in KIT. Only 17% of the deletion mutations were detected, because the panel was not designed for detecting these. A substitution in FBX4 exon 1 (S8R), representing a germline single-nucleotide polymorphism, was observed in a case with KIT exon 11 missense mutation. No other mutations were identified, including in the six wild-type GISTs. Conclusions: Our results indicate that mutations other than KIT or PDGFRA are rare in GISTs. Although multiplex mutation screening by mass spectrometry detected missense mutations accurately, it is not sufficient to screen mutations because deletion mutations are common in GISTs.
KW - Gastrointestinal stromal tumour
KW - High-Throughput screening
KW - Mass spectrometry
KW - Mutation
KW - Sequencin
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U2 - 10.1097/PAT.0b013e3283559c45
DO - 10.1097/PAT.0b013e3283559c45
M3 - Article
C2 - 22777070
AN - SCOPUS:84866943676
SN - 0031-3025
VL - 44
SP - 460
EP - 464
JO - Pathology
JF - Pathology
IS - 5
ER -