用于稳定同位素分辨代谢组学的培养哺乳动物细胞代谢标记：组织培养和样品提取的实际操作
Metabolic Labeling of Cultured Mammalian Cells for Stable Isotope-Resolved Metabolomics: Practical Aspects of Tissue Culture and Sample Extraction.
作者信息
Crooks Daniel R, Fan Teresa W-M, Linehan W Marston
机构信息
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.
出版信息
Methods Mol Biol. 2019;1928:1-27. doi: 10.1007/978-1-4939-9027-6_1.
Abstract
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.
摘要
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。
相似文献
1
Metabolic Labeling of Cultured Mammalian Cells for Stable Isotope-Resolved Metabolomics: Practical Aspects of Tissue Culture and Sample Extraction.用于稳定同位素分辨代谢组学的培养哺乳动物细胞代谢标记：组织培养和样品提取的实际操作
Methods Mol Biol. 2019;1928:1-27. doi: 10.1007/978-1-4939-9027-6_1.
3
Stable Isotope-Resolved Metabolomics by NMR.基于核磁共振的稳定同位素分辨代谢组学
Methods Mol Biol. 2019;2037:151-168. doi: 10.1007/978-1-4939-9690-2_9.
4
NMR-Based Stable Isotope Tracing of Cancer Metabolism.基于 NMR 的癌症代谢稳定同位素示踪。
Methods Mol Biol. 2025;2855:457-504. doi: 10.1007/978-1-0716-4116-3_26.
5
Stable isotope-resolved metabolomics and applications for drug development.稳定同位素分辨代谢组学及其在药物开发中的应用。
Pharmacol Ther. 2012 Mar;133(3):366-91. doi: 10.1016/j.pharmthera.2011.12.007. Epub 2011 Dec 23.
6
Tracer-Based Cancer Metabolomic Analysis.基于示踪剂的癌症代谢组学分析。
Adv Exp Med Biol. 2021;1280:115-130. doi: 10.1007/978-3-030-51652-9_8.
7
Chloroformate derivatization for tracing the fate of Amino acids in cells and tissues by multiple stable isotope resolved metabolomics (mSIRM).氯甲酸酯衍生化法用于通过多重稳定同位素分辨代谢组学（mSIRM）追踪细胞和组织中氨基酸的命运。
Anal Chim Acta. 2017 Jul 11;976:63-73. doi: 10.1016/j.aca.2017.04.014. Epub 2017 Apr 10.
8
Probing the metabolic phenotype of breast cancer cells by multiple tracer stable isotope resolved metabolomics.采用多种示踪稳定同位素解析代谢组学技术探究乳腺癌细胞的代谢表型。
Metab Eng. 2017 Sep;43(Pt B):125-136. doi: 10.1016/j.ymben.2017.01.010. Epub 2017 Feb 2.
9
Stable isotope-resolved metabolomics analyses of metabolic phenotypes reveal variable glutamine metabolism in different patient-derived models of non-small cell lung cancer from a single patient.稳定同位素分辨代谢组学分析代谢表型揭示了来自单一患者的不同非小细胞肺癌患者衍生模型中可变的谷氨酰胺代谢。
Metabolomics. 2024 Jul 27;20(4):87. doi: 10.1007/s11306-024-02126-x.
10
Integration of flux measurements and pharmacological controls to optimize stable isotope-resolved metabolomics workflows and interpretation.通量测量与药理学控制的整合，以优化稳定同位素解析代谢组学工作流程和解释。
Sci Rep. 2019 Sep 23;9(1):13705. doi: 10.1038/s41598-019-50183-3.
引用本文的文献
1
Understanding the Metabolic Effects of Surgically Induced Renal Ischemia in Humans: A Temporal Approach.了解手术诱导的人类肾缺血的代谢效应：一种时间性方法。
Metabolites. 2025 Jul 8;15(7):462. doi: 10.3390/metabo15070462.
2
Metabolic Pathway Tracing for NAD Synthesis and Consumption.用于NAD合成与消耗的代谢途径追踪
Methods Mol Biol. 2025;2925:203-222. doi: 10.1007/978-1-0716-4534-5_14.
3
Targeting metabolic vulnerability by combining NAMPT inhibitors and disulfiram for treatment of recurrent ovarian cancer.联合使用烟酰胺磷酸核糖转移酶（NAMPT）抑制剂和双硫仑靶向代谢脆弱性以治疗复发性卵巢癌。
Cell Death Dis. 2025 Apr 25;16(1):342. doi: 10.1038/s41419-025-07672-3.
4
Differences in glycolytic metabolism between tissue-resident alveolar macrophages and recruited lung macrophages.组织驻留肺泡巨噬细胞和募集的肺巨噬细胞之间糖酵解代谢的差异。
Front Immunol. 2025 Feb 28;16:1535796. doi: 10.3389/fimmu.2025.1535796. eCollection 2025.
5
NMR-Based Stable Isotope Tracing of Cancer Metabolism.基于 NMR 的癌症代谢稳定同位素示踪。
Methods Mol Biol. 2025;2855:457-504. doi: 10.1007/978-1-0716-4116-3_26.
6
Stable Isotope Tracing Analysis in Cancer Research: Advancements and Challenges in Identifying Dysregulated Cancer Metabolism and Treatment Strategies.癌症研究中的稳定同位素示踪分析：识别失调的癌症代谢及治疗策略方面的进展与挑战
Metabolites. 2024 May 31;14(6):318. doi: 10.3390/metabo14060318.
7
Determination of fatty acid uptake and desaturase activity in mammalian cells by NMR-based stable isotope tracing.基于 NMR 稳定同位素示踪技术测定哺乳动物细胞中脂肪酸的摄取和去饱和酶活性。
Anal Chim Acta. 2024 May 15;1303:342511. doi: 10.1016/j.aca.2024.342511. Epub 2024 Mar 21.
8
Targeting mutant p53-R248W reactivates WT p53 function and alters the onco-metabolic profile.靶向突变型p53-R248W可恢复野生型p53功能并改变肿瘤代谢谱。
Front Oncol. 2023 Jan 11;12:1094210. doi: 10.3389/fonc.2022.1094210. eCollection 2022.
9
Polarization and β-Glucan Reprogram Immunomodulatory Metabolism in Human Macrophages and Ex Vivo in Human Lung Cancer Tissues.极化和β-葡聚糖重塑人巨噬细胞的免疫调节代谢，并在人肺癌组织中进行离体研究。
J Immunol. 2022 Nov 1;209(9):1674-1690. doi: 10.4049/jimmunol.2200178. Epub 2022 Sep 23.
10
Simple Esterification of [1-C]-Alpha-Ketoglutarate Enhances Membrane Permeability and Allows for Noninvasive Tracing of Glutamate and Glutamine Production.[1-C]-α-酮戊二酸的简单酯化增强了膜通透性，并允许非侵入性追踪谷氨酸和谷氨酰胺的产生。
ACS Chem Biol. 2021 Nov 19;16(11):2144-2150. doi: 10.1021/acschembio.1c00561. Epub 2021 Sep 23.
本文引用的文献
1
Metabolomics and Isotope Tracing.代谢组学和同位素示踪。
Cell. 2018 May 3;173(4):822-837. doi: 10.1016/j.cell.2018.03.055.
2
Acute loss of iron-sulfur clusters results in metabolic reprogramming and generation of lipid droplets in mammalian cells.急性铁硫簇缺失导致哺乳动物细胞代谢重编程和脂滴生成。
J Biol Chem. 2018 May 25;293(21):8297-8311. doi: 10.1074/jbc.RA118.001885. Epub 2018 Mar 9.
3
Quantitative profiling of carbonyl metabolites directly in crude biological extracts using chemoselective tagging and nanoESI-FTMS.采用化学选择性标记和纳升电喷雾傅里叶变换质谱法直接对粗生物提取物中的羰基代谢物进行定量分析。
Analyst. 2017 Dec 18;143(1):311-322. doi: 10.1039/c7an01256j.
4
Noninvasive liquid diet delivery of stable isotopes into mouse models for deep metabolic network tracing.非侵入性液体饮食递送来对小鼠模型进行深入代谢网络示踪的稳定同位素。
Nat Commun. 2017 Nov 21;8(1):1646. doi: 10.1038/s41467-017-01518-z.
5
Glucose feeds the TCA cycle via circulating lactate.葡萄糖通过循环的乳酸为三羧酸循环提供能量。
Nature. 2017 Nov 2;551(7678):115-118. doi: 10.1038/nature24057. Epub 2017 Oct 18.
6
Lactate Metabolism in Human Lung Tumors.人类肺部肿瘤中的乳酸代谢
Cell. 2017 Oct 5;171(2):358-371.e9. doi: 10.1016/j.cell.2017.09.019.
7
Exploring cancer metabolism using stable isotope-resolved metabolomics (SIRM).利用稳定同位素分辨代谢组学（SIRM）探索癌症代谢。
J Biol Chem. 2017 Jul 14;292(28):11601-11609. doi: 10.1074/jbc.R117.776054. Epub 2017 Jun 7.
8
Chloroformate derivatization for tracing the fate of Amino acids in cells and tissues by multiple stable isotope resolved metabolomics (mSIRM).氯甲酸酯衍生化法用于通过多重稳定同位素分辨代谢组学（mSIRM）追踪细胞和组织中氨基酸的命运。
Anal Chim Acta. 2017 Jul 11;976:63-73. doi: 10.1016/j.aca.2017.04.014. Epub 2017 Apr 10.
9
CPS1 maintains pyrimidine pools and DNA synthesis in KRAS/LKB1-mutant lung cancer cells.CPS1维持KRAS/LKB1突变型肺癌细胞中的嘧啶库和DNA合成。
Nature. 2017 Jun 1;546(7656):168-172. doi: 10.1038/nature22359. Epub 2017 May 24.
10
Physiologic Medium Rewires Cellular Metabolism and Reveals Uric Acid as an Endogenous Inhibitor of UMP Synthase.生理介质重塑细胞代谢并揭示尿酸是尿苷一磷酸合酶的内源性抑制剂。
Cell. 2017 Apr 6;169(2):258-272.e17. doi: 10.1016/j.cell.2017.03.023.
Zotero 插件