优化的用于稳定同位素示踪和荷人表皮生长因子受体 2（HER2）阳性乳腺癌脑转移小鼠的稳态代谢组学的方案。

Optimized protocol for stable isotope tracing and steady-state metabolomics in mouse HER2+ breast cancer brain metastasis.

机构信息

Department of Pathology, UT Southwestern Medical Center, Dallas, TX 75390, USA.

Harold C. Simmons Comprehensive Cancer Center, UT Southwestern Medical Center, Dallas, TX, USA.

出版信息

STAR Protoc. 2022 Apr 22;3(2):101345. doi: 10.1016/j.xpro.2022.101345. eCollection 2022 Jun 17.

DOI:10.1016/j.xpro.2022.101345

PMID:35496802

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9048131/

Abstract

Analyzing the metabolic dependencies of tumor cells is vital for cancer diagnosis and treatment. Here, we describe a protocol for C-stable glucose and glutamine isotope tracing in mice HER2+ breast cancer brain metastatic lesions. We describe how to inject cancer cells intracardially to generate brain metastatic lesions in mice. We then detail how to perform C-stable isotope infusion in mice with established brain metastasis. Finally, we outline steps for sample collection, processing for metabolite extraction, and analyzing mass spectrometry data. For complete details on the use and execution of this protocol, please refer to Parida et al. (2022).

摘要

分析肿瘤细胞的代谢依赖性对于癌症的诊断和治疗至关重要。在这里，我们描述了一种在小鼠 HER2+乳腺癌脑转移病灶中进行 C-稳定葡萄糖和谷氨酰胺同位素示踪的方案。我们描述了如何通过心内注射将癌细胞注入小鼠体内以生成脑转移病灶。然后，我们详细介绍了如何在已建立脑转移的小鼠中进行 C-稳定同位素输注。最后，我们概述了样本收集、代谢物提取的处理以及质谱数据分析的步骤。有关该方案使用和执行的完整详细信息，请参见 Parida 等人（2022 年）。