Iorio Egidio, Ricci Alessandro, Pisanu Maria Elena, Bagnoli Marina, Podo Franca, Canevari Silvana
Department of Cell Biology and Neurosciences, Istituto Superiore di Sanità, Rome, Italy.
Methods Mol Biol. 2013;1049:255-70. doi: 10.1007/978-1-62703-547-7_19.
The revised version of cancer hallmarks, depicting the biological properties acquired during tumor development and progression, includes the capability to modify or reprogram cellular metabolism. High-resolution multinuclear magnetic resonance spectroscopy (MRS) provides noninvasive means of monitoring metabolites that play a central role in several pathways, measuring the rates at which reactions within the pathways take place, and investigating how these rates are controlled in such a way that a metabolic precursor and cofactors are provided according to the demand. Here we describe comprehensive methods for assaying the activity of key enzymes involved in the phosphatidylcholine metabolic pathways responsible for phosphocholine accumulation in ovarian cancer cells using high-resolution (1)H and (31)P MRS analyses of cell lysates or cytosolic preparations.
癌症特征的修订版描述了肿瘤发生和发展过程中获得的生物学特性,其中包括改变或重编程细胞代谢的能力。高分辨率多核磁共振波谱(MRS)提供了一种非侵入性手段,可用于监测在多个途径中起核心作用的代谢物,测量途径内反应发生的速率,并研究这些速率是如何被控制的,以便根据需求提供代谢前体和辅因子。在这里,我们描述了使用细胞裂解物或胞质制剂的高分辨率¹H和³¹P MRS分析,来测定参与卵巢癌细胞中磷酸胆碱积累的磷脂酰胆碱代谢途径中关键酶活性的综合方法。