Murthy Divya, Vernucci Enza, Goode Gennifer, Abrego Jaime, Singh Pankaj K
The Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha, NE, USA.
Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, NE, USA.
Methods Mol Biol. 2018;1742:81-93. doi: 10.1007/978-1-4939-7665-2_8.
Hypoxia is frequently observed in human cancers and induces global metabolic reprogramming that includes an increase in glucose uptake and glycolysis, alterations in NAD(P)H/NAD(P) and intracellular ATP levels, and increased utilization of glutamine as the major precursor for fatty acid synthesis. In this chapter, we describe in detail various physiological assays that have been adopted to study the metabolic shift propagated by exposure to hypoxic conditions in pancreatic cell culture model that includes glucose uptake, glutamine uptake, and lactate release by pancreatic cancer cell lines. We have also elaborated the assays to evaluate the ratio of NAD(P)H/NAD(P) and intracellular ATP estimation using the commercially available kit to assess the metabolic state of cancer cells.
缺氧在人类癌症中经常被观察到，并诱导整体代谢重编程，包括葡萄糖摄取和糖酵解增加、NAD(P)H/NAD(P)和细胞内ATP水平改变，以及谷氨酰胺作为脂肪酸合成主要前体的利用率增加。在本章中，我们详细描述了各种生理学检测方法，这些方法已被用于研究在胰腺细胞培养模型中暴露于缺氧条件下所引发的代谢转变，包括胰腺癌细胞系的葡萄糖摄取、谷氨酰胺摄取和乳酸释放。我们还阐述了使用市售试剂盒评估NAD(P)H/NAD(P)比值和细胞内ATP估计值以评估癌细胞代谢状态的检测方法。
