Chung Y-L, Leach M O, Eykyn T R
Cancer Research UK Cancer Imaging Centre, The Institute of Cancer Research, London, United Kingdom.
Cancer Research UK Cancer Imaging Centre, The Institute of Cancer Research, London, United Kingdom.
Methods Enzymol. 2017;588:133-153. doi: 10.1016/bs.mie.2016.09.078. Epub 2016 Nov 17.
Cancer cells undergoing starvation- and treatment-induced autophagy were found to exhibit reduced intracellular lactate, reduced rates of steady-state lactate excretion and reduced real-time pyruvate-lactate exchange rates, indicating that glycolytic metabolism was altered in autophagic cells. In this chapter, we describe the technical details of the use of H-magnetic resonance spectroscopy (MRS) to measure endogenous cellular concentrations of lactate and glucose in autophagic cells and tissues, how to measure the rate of steady-state lactate excretion and glucose uptake by H-MRS in autophagic cells, and details of the real-time measurement of [1-C] pyruvate to lactate exchange in autophagic cells by C-MRS-DNP (dynamic nuclear polarization).
研究发现,经历饥饿和治疗诱导自噬的癌细胞表现出细胞内乳酸减少、稳态乳酸排泄率降低以及实时丙酮酸-乳酸交换率降低,这表明自噬细胞中的糖酵解代谢发生了改变。在本章中,我们描述了使用氢磁共振波谱(MRS)测量自噬细胞和组织中内源性细胞乳酸和葡萄糖浓度的技术细节,如何通过氢MRS测量自噬细胞中稳态乳酸排泄率和葡萄糖摄取率,以及通过碳MRS-动态核极化(C-MRS-DNP)实时测量自噬细胞中[1-碳]丙酮酸向乳酸交换的细节。