Absolute quantitation of Met using mass spectrometry for clinical application: assay precision, stability, and correlation with MET gene amplification in FFPE tumor tissue
| Autor: | Shu-Yuan Xiao, Brittany Rambo, Fabiola Cecchi, Jon Burrows, Daniel V.T. Catenacci, Wei-Li Liao, Lei Zhao, David B. Krizman, Kathleen Bengali, Peng Xu, Theodore Karrison, Jamar Uzzell, Marlene Darfler, Les Henderson, Todd Hembrough, Sheeno Thyparambil, John Hart, Donald P. Bottaro, Timothy D. Veenstra |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2014 |
| Předmět: |
Male
Pathology Esophageal Neoplasms Cancer Treatment lcsh:Medicine Biochemistry Mass Spectrometry Metastasis law.invention Analytical Chemistry 0302 clinical medicine Secondary Ion Mass Spectrometry Spectrum Analysis Techniques law Gene duplication Gastrointestinal Cancers Medicine and Health Sciences Copy-number variation Clinical Trials (Cancer Treatment) lcsh:Science 0303 health sciences Multidisciplinary medicine.diagnostic_test Cancer Risk Factors Proto-Oncogene Proteins c-met Immunohistochemistry Chemistry Oncology 030220 oncology & carcinogenesis Physical Sciences Recombinant DNA Female Research Article medicine.medical_specialty Esophageal Cancer Genetic Causes of Cancer Gastroenterology and Hepatology Biology Research and Analysis Methods 03 medical and health sciences Stomach Neoplasms Biopsy Gastrointestinal Tumors medicine Humans Molecular Biology Techniques Molecular Biology Immunohistochemistry Techniques 030304 developmental biology lcsh:R Gene Amplification Biology and Life Sciences Cancers and Neoplasms medicine.disease Gene expression profiling Histochemistry and Cytochemistry Techniques Gastric Cancer Cancer research lcsh:Q Biomarkers Fluorescence in situ hybridization |
| Zdroj: | PLoS ONE, Vol 9, Iss 7, p e100586 (2014) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | Background Overexpression of Met tyrosine kinase receptor is associated with poor prognosis. Overexpression, and particularly MET amplification, are predictive of response to Met-specific therapy in preclinical models. Immunohistochemistry (IHC) of formalin-fixed paraffin-embedded (FFPE) tissues is currently used to select for ‘high Met’ expressing tumors for Met inhibitor trials. IHC suffers from antibody non-specificity, lack of quantitative resolution, and, when quantifying multiple proteins, inefficient use of scarce tissue. Methods After describing the development of the Liquid-Tissue-Selected Reaction Monitoring-mass spectrometry (LT-SRM-MS) Met assay, we evaluated the expression level of Met in 130 FFPE gastroesophageal cancer (GEC) tissues. We assessed the correlation of SRM Met expression to IHC and mean MET gene copy number (GCN)/nucleus or MET/CEP7 ratio by fluorescence in situ hybridization (FISH). Results Proteomic mapping of recombinant Met identified 418TEFTTALQR426 as the optimal SRM peptide. Limits of detection (LOD) and quantitation (LOQ) for this peptide were 150 and 200 amol/µg tumor protein, respectively. The assay demonstrated excellent precision and temporal stability of measurements in serial sections analyzed one year apart. Expression levels of 130 GEC tissues ranged ( |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|