Liu Ning Qing, Dekker Lennard J M, Van Duijn Martijn M, Umar Arzu
Department of Medical Oncology, Laboratory of Breast Cancer Genomics and Proteomics, Erasmus University Medical Center, Erasmus MC Cancer Institute, Wytemaweg 80, Be-401, 2040, 3000, CA, Rotterdam, The Netherlands.
Methods Mol Biol. 2014;1156:307-22. doi: 10.1007/978-1-4939-0685-7_21.
Mass spectrometry-based proteomics facilitates high-throughput discovery of protein markers for diagnosis and treatment of breast cancer patients. Hundreds of putative prognostic and predictive markers are being identified every year, but only a very small proportion of them can be validated as clinically relevant markers. A quantitative and cost-efficient verification method is highly desirable to pick up real "nuggets" from the "sand." To fulfill these criteria, we previously introduced a stable isotope labeling by amino acids in cell culture (SILAC)-based selected reaction monitoring (SRM) approach for studying breast cancer-related protein markers. Here we describe a hands-on protocol of using this SILAC-SRM method for verification of breast cancer-related markers, which can also be used for verification of protein markers in other types of solid tumor tissues.
基于质谱的蛋白质组学有助于高通量发现用于乳腺癌患者诊断和治疗的蛋白质标志物。每年都有数百种假定的预后和预测标志物被鉴定出来，但其中只有极小一部分能够被验证为具有临床相关性的标志物。非常需要一种定量且经济高效的验证方法，以便从“沙子”中筛选出真正的“金子”。为满足这些标准，我们之前引入了一种基于细胞培养中氨基酸稳定同位素标记（SILAC）的选择反应监测（SRM）方法来研究乳腺癌相关蛋白质标志物。在此，我们描述了一种使用这种SILAC-SRM方法验证乳腺癌相关标志物的实用方案，该方案也可用于验证其他类型实体瘤组织中的蛋白质标志物。
