Pietzke Matthias, Kempa Stefan
Berlin Institute for Medical Systems Biology at the MDC Berlin-Buch, Berlin, Germany.
Berlin Institute for Medical Systems Biology at the MDC Berlin-Buch, Berlin, Germany.
Methods Enzymol. 2014;543:179-98. doi: 10.1016/B978-0-12-801329-8.00009-X.
Metabolic reprogramming is a key step in oncogenic transformation, and it involves alterations in both bioenergetic and anabolic metabolism. Sustained by these metabolic alterations, malignant cells acquire the ability to re-enter the cell cycle and proliferate. The so-called central carbon metabolism (CCM) is the ultimate source for energy and building blocks enabling cellular growth and proliferation. The time-resolved monitoring of the conversion of stable isotope-labeled metabolites provides profound insights into the metabolic dynamics of malignant cells and enables the tracking of individual carbon routes within the CCM. Specifically, the analysis of isotope incorporation rates within short time frames by means of pulsed stable isotope-resolved metabolomics (pSIRM) can be used to determine the dynamics of glycolysis and glutaminolysis-two metabolic circuitries that are often deregulated in malignant cells. Here, we detail a pSIRM-based method that can be applied to the study of metabolic alteration in cultured cancer cells.
代谢重编程是致癌转化的关键步骤，它涉及生物能量代谢和合成代谢的改变。在这些代谢改变的支持下，恶性细胞获得了重新进入细胞周期并增殖的能力。所谓的中心碳代谢（CCM）是能量和构建细胞生长与增殖所需物质的最终来源。对稳定同位素标记代谢物转化的时间分辨监测，能深入了解恶性细胞的代谢动态，并追踪CCM内的各个碳途径。具体而言，通过脉冲稳定同位素分辨代谢组学（pSIRM）在短时间内分析同位素掺入率，可用于确定糖酵解和谷氨酰胺分解的动态——这两种代谢途径在恶性细胞中常常失调。在此，我们详细介绍一种基于pSIRM的方法，该方法可应用于研究培养的癌细胞中的代谢改变。
