Crooks Daniel R, Fan Teresa W-M, Linehan W Marston
Urologic Oncology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA.
Department of Toxicology and Cancer Biology, Center for Environmental and Systems Biochemistry, Markey Cancer Center, and University of Kentucky, Lexington, KY, USA.
Methods Mol Biol. 2019;1928:1-27. doi: 10.1007/978-1-4939-9027-6_1.
Stable isotope-resolved metabolomics (SIRM) methods are used increasingly by cancer researchers to probe metabolic pathways and identify vulnerabilities in cancer cells. Analytical and computational advances are being made constantly, but tissue culture and sample extraction procedures are often variable and not elaborated in the literature. This chapter discusses basic aspects of tissue culture practices as they relate to the use of stable isotope tracers and provides a detailed metabolic labeling and metabolite extraction procedure designed to maximize the amount of information that can be obtained from a single tracer experiment.
癌症研究人员越来越多地使用稳定同位素分辨代谢组学（SIRM）方法来探究代谢途径并识别癌细胞中的脆弱点。分析和计算方面不断取得进展，但组织培养和样品提取程序往往各不相同，且文献中并未详细阐述。本章讨论了与稳定同位素示踪剂使用相关的组织培养实践的基本方面，并提供了详细的代谢标记和代谢物提取程序，旨在最大限度地从单个示踪剂实验中获取信息量。
