利用 Seahorse 技术分析黑素瘤细胞耗氧量和细胞外酸化率。
Analyzing Melanoma Cell Oxygen Consumption and Extracellular Acidification Rates Using Seahorse Technology.
机构信息
Tumor Microenvironment Center, UPMC Hillman Cancer Center, University of Pittsburgh, Pittsburgh, PA, USA.
Department of Immunology, University of Pittsburgh, Pittsburgh, PA, USA.
出版信息
Methods Mol Biol. 2021;2265:81-89. doi: 10.1007/978-1-0716-1205-7_6.
Abstract
Cancer cells have deregulated metabolism that can contribute to the unique metabolic makeup of the tumor microenvironment. This can be variable between patients, and it is important to understand these differences since they potentially can affect therapy response. Here we discuss a method of processing and assaying metabolism from direct ex vivo murine and human tumor samples using seahorse extracellular flux analysis. This provides real-time profiling of oxidative versus glycolytic metabolism and can help infer the metabolic status of the tumor microenvironment.
摘要
癌细胞的代谢失调可能导致肿瘤微环境的独特代谢构成。这种代谢失调在患者之间可能存在差异，了解这些差异非常重要，因为它们可能会影响治疗反应。在这里，我们讨论了一种使用 Seahorse 细胞外通量分析直接从体外培养的鼠和人肿瘤样本中处理和分析代谢的方法。该方法可实时分析氧化代谢与糖酵解代谢，并有助于推断肿瘤微环境的代谢状态。
相似文献
1
Analyzing Melanoma Cell Oxygen Consumption and Extracellular Acidification Rates Using Seahorse Technology.利用 Seahorse 技术分析黑素瘤细胞耗氧量和细胞外酸化率。
Methods Mol Biol. 2021;2265:81-89. doi: 10.1007/978-1-0716-1205-7_6.
2
Extracellular Flux Assays to Determine Oxidative Phosphorylation and Glycolysis in Chronic Lymphocytic Leukemia Cells.用于测定慢性淋巴细胞白血病细胞中氧化磷酸化和糖酵解的细胞外通量分析
Methods Mol Biol. 2019;1881:121-128. doi: 10.1007/978-1-4939-8876-1_10.
3
Studying the Metabolism of Epithelial-Mesenchymal Plasticity Using the Seahorse XFe96 Extracellular Flux Analyzer.使用 Seahorse XFe96 细胞外通量分析仪研究上皮-间充质可塑性的代谢。
Methods Mol Biol. 2021;2179:327-340. doi: 10.1007/978-1-0716-0779-4_25.
4
Evaluating the Role of RARβ Signaling on Cellular Metabolism in Melanoma Using the Seahorse XF Analyzer.使用海马XF分析仪评估RARβ信号在黑色素瘤细胞代谢中的作用。
Methods Mol Biol. 2019;2019:171-180. doi: 10.1007/978-1-4939-9585-1_12.
5
Determining Compartment-Specific Metabolic Fluxes.确定特定隔室的代谢通量。
Methods Mol Biol. 2019;1862:137-149. doi: 10.1007/978-1-4939-8769-6_10.
6
Measurement of mitochondrial respiration in the murine retina using a Seahorse extracellular flux analyzer.使用 Seahorse 细胞外通量分析仪测量鼠视网膜中的线粒体呼吸。
STAR Protoc. 2021 May 12;2(2):100533. doi: 10.1016/j.xpro.2021.100533. eCollection 2021 Jun 18.
7
Bioenergetic analysis of intact mammalian cells using the Seahorse XF24 Extracellular Flux analyzer and a luciferase ATP assay.使用海马XF24细胞外通量分析仪和荧光素酶ATP检测法对完整哺乳动物细胞进行生物能量分析。
Methods Mol Biol. 2014;1105:589-602. doi: 10.1007/978-1-62703-739-6_40.
8
Determining Macrophage Polarization upon Metabolic Perturbation.代谢扰动时巨噬细胞极化的测定
Methods Mol Biol. 2019;1862:173-186. doi: 10.1007/978-1-4939-8769-6_13.
9
Using Seahorse Machine to Measure OCR and ECAR in Cancer Cells.使用海马体分析仪测量癌细胞中的氧消耗率（OCR）和细胞外酸化率（ECAR）。
Methods Mol Biol. 2019;1928:353-363. doi: 10.1007/978-1-4939-9027-6_18.
10
Extracellular flux assay (Seahorse assay): Diverse applications in metabolic research across biological disciplines.细胞外通量分析（ Seahorse 分析）：在跨生物学科的代谢研究中的多种应用。
Mol Cells. 2024 Aug;47(8):100095. doi: 10.1016/j.mocell.2024.100095. Epub 2024 Jul 18.
本文引用的文献
1
Tumor cell oxidative metabolism as a barrier to PD-1 blockade immunotherapy in melanoma.肿瘤细胞氧化代谢作为黑色素瘤 PD-1 阻断免疫治疗的障碍。
JCI Insight. 2019 Mar 7;4(5). doi: 10.1172/jci.insight.124989.
2
Metabolic Switch in the Tumor Microenvironment Determines Immune Responses to Anti-cancer Therapy.肿瘤微环境中的代谢转换决定了对抗癌治疗的免疫反应。
Front Oncol. 2018 Aug 13;8:284. doi: 10.3389/fonc.2018.00284. eCollection 2018.
3
4-1BB costimulation induces T cell mitochondrial function and biogenesis enabling cancer immunotherapeutic responses.4-1BB 共刺激诱导 T 细胞线粒体功能和生物发生，从而实现癌症免疫治疗反应。
J Exp Med. 2018 Apr 2;215(4):1091-1100. doi: 10.1084/jem.20171068. Epub 2018 Mar 6.
4
Early TCR Signaling Induces Rapid Aerobic Glycolysis Enabling Distinct Acute T Cell Effector Functions.早期 TCR 信号诱导快速有氧糖酵解，从而实现独特的急性 T 细胞效应功能。
Cell Rep. 2018 Feb 6;22(6):1509-1521. doi: 10.1016/j.celrep.2018.01.040.
5
Efficacy of PD-1 Blockade Is Potentiated by Metformin-Induced Reduction of Tumor Hypoxia.PD-1 阻断疗法的疗效通过二甲双胍诱导的肿瘤缺氧减轻而增强。
Cancer Immunol Res. 2017 Jan;5(1):9-16. doi: 10.1158/2326-6066.CIR-16-0103. Epub 2016 Dec 9.
6
Tumor Microenvironment Metabolism: A New Checkpoint for Anti-Tumor Immunity.肿瘤微环境代谢：抗肿瘤免疫的新检查点
Vaccines (Basel). 2016 Dec 6;4(4):46. doi: 10.3390/vaccines4040046.
7
The Tumor Microenvironment Represses T Cell Mitochondrial Biogenesis to Drive Intratumoral T Cell Metabolic Insufficiency and Dysfunction.肿瘤微环境抑制T细胞线粒体生物合成，以驱动肿瘤内T细胞代谢不足和功能障碍。
Immunity. 2016 Aug 16;45(2):374-88. doi: 10.1016/j.immuni.2016.07.009. Epub 2016 Aug 2.
8
Rapid analysis of glycolytic and oxidative substrate flux of cancer cells in a microplate.微孔板中癌细胞糖酵解和氧化底物通量的快速分析
PLoS One. 2014 Oct 31;9(10):e109916. doi: 10.1371/journal.pone.0109916. eCollection 2014.
9
Mitochondrially targeted compounds and their impact on cellular bioenergetics.线粒体靶向化合物及其对细胞生物能量学的影响。
Redox Biol. 2013;1(1):86-93. doi: 10.1016/j.redox.2012.11.009.
10
Understanding the Warburg effect: the metabolic requirements of cell proliferation.理解瓦伯格效应：细胞增殖的代谢需求。
Science. 2009 May 22;324(5930):1029-33. doi: 10.1126/science.1160809.
Zotero 插件