Klawitter J, Anderson N, Klawitter J, Christians U, Leibfritz D, Eckhardt S G, Serkova N J
Department of Anesthesiology, University of Colorado, Denver, CO 80045, USA.
Br J Cancer. 2009 Mar 24;100(6):923-31. doi: 10.1038/sj.bjc.6604946. Epub 2009 Mar 3.
The goal of this study was to evaluate the time course of metabolic changes in leukaemia cells treated with the Bcr-Abl tyrosine kinase inhibitor imatinib. Human Bcr-Abl(+) K562 cells were incubated with imatinib in a dose-escalating manner (starting at 0.1 microM with a weekly increase of 0.1 microM imatinib) for up to 5 weeks. Nuclear magnetic resonance spectroscopy and liquid-chromatography mass spectrometry were performed to assess a global metabolic profile, including glucose metabolism, energy state, lipid metabolism and drug uptake, after incubation with imatinib. Initially, imatinib treatment completely inhibited the activity of Bcr-Abl tyrosine kinase, followed by the inhibition of cell glycolytic activity and glucose uptake. This was accompanied by the increased mitochondrial activity and energy production. With escalating imatinib doses, the process of cell death rapidly progressed. Phosphocreatine and NAD(+) concentrations began to decrease, and mitochondrial activity, as well as the glycolysis rate, was further reduced. Subsequently, the synthesis of lipids as necessary membrane precursors for apoptotic bodies was accelerated. The concentrations of the Kennedy pathway intermediates, phosphocholine and phosphatidylcholine, were reduced. After 4 weeks of exposure to imatinib, the secondary necrosis associated with decrease in the mitochondrial and glycolytic activity occurred and was followed by a shutdown of energy production and cell death. In conclusion, monitoring of metabolic changes in cells exposed to novel signal transduction modulators supplements molecular findings and provides further mechanistic insights into longitudinal changes of the mitochondrial and glycolytic pathways of oncogenesis.
本研究的目的是评估用Bcr-Abl酪氨酸激酶抑制剂伊马替尼治疗的白血病细胞代谢变化的时间进程。将人Bcr-Abl(+) K562细胞与伊马替尼以剂量递增的方式（从0.1微摩尔开始，每周增加0.1微摩尔伊马替尼）孵育长达5周。在用伊马替尼孵育后，进行核磁共振波谱和液相色谱质谱分析以评估整体代谢概况，包括葡萄糖代谢、能量状态、脂质代谢和药物摄取。最初，伊马替尼治疗完全抑制了Bcr-Abl酪氨酸激酶的活性，随后抑制了细胞糖酵解活性和葡萄糖摄取。这伴随着线粒体活性和能量产生的增加。随着伊马替尼剂量的增加，细胞死亡过程迅速进展。磷酸肌酸和NAD(+)浓度开始降低，线粒体活性以及糖酵解速率进一步降低。随后，作为凋亡小体必要膜前体的脂质合成加速。肯尼迪途径中间体磷酸胆碱和磷脂酰胆碱的浓度降低。在暴露于伊马替尼4周后，发生了与线粒体和糖酵解活性降低相关的继发性坏死，随后能量产生停止和细胞死亡。总之，监测暴露于新型信号转导调节剂的细胞中的代谢变化补充了分子研究结果，并为肿瘤发生的线粒体和糖酵解途径的纵向变化提供了进一步的机制见解。
