Metabolic characteristics of imatinib resistance in chronic myeloid leukaemia cells
Metabolic characteristics of imatinib resistance in chronic myeloid leukaemia cells
复制标题
DOI：
10.1111/j.1476-5381.2009.00345.x
复制
发表时间：
2009-09-01
影响因子：
7.3
通讯作者：
Serkova, Natalie J.
中科院分区：
文献类型：
--
作者：
Klawitter, Jelena;Kominsky, Douglas J.;Serkova, Natalie J.
背景和目的：早期发现耐药发展对于以伊马替尼为基础治疗慢性髓性白血病（CML）患者至关重要。我们的目的是区分细胞对伊马替尼耐药的代谢标志物。实验方法：通过核磁共振波谱分析两种人类伊马替尼敏感CML细胞系LAMA84-s和K562-s，以及它们的耐药对应细胞系LAMA84-r和K562-r（均对1 μ M伊马替尼耐药）和K562-r (5 μ M)，以评估整体代谢谱，包括能量状态、葡萄糖和磷脂代谢。关键结果：我们通过Western blotting和流式细胞术发现耐药克隆中Bcr-Abl酪氨酸激酶和多药耐药p糖蛋白水平不一致。另一方面，磷脂代谢和乳酸生成高度预测细胞对伊马替尼的反应。如前所述，敏感细胞在1 μ M伊马替尼处理24 h后，糖酵解活性（乳酸）和磷脂合成（磷脂胆碱）明显降低，磷脂分解代谢（甘油磷脂胆碱）增加，这与抑制细胞增殖和诱导细胞凋亡有关。与它们的敏感对应体相比，K562-r、K562-r和LAMA84-r在1 μ M （K562-r和LAMA84-r）和5 μ M （K562-r）伊马替尼的存在下，磷脂合成和糖酵解乳酸产量保持增加。结论和意义：可以在耐药克隆中发现早期检测伊马替尼耐药的特定代谢标志物，包括糖酵解活性和磷脂转化的增加。一旦在人类分离的白细胞中得到验证，它们可用于监测CML患者对治疗的反应性。
Background and purpose:Early detection of resistance development is crucial for imatinib-based treatment in chronic myeloid leukaemia (CML) patients. We aimed to distinguish metabolic markers of cell resistance to imatinib.Experimental approach:Two human imatinib-sensitive CML cell lines: LAMA84-s and K562-s, and their resistant counterparts: LAMA84-r and K562-r (both resistant to 1 mu M imatinib), and K562-R (5 mu M) were analysed by nuclear magnetic resonance spectroscopy to assess global metabolic profiling, including energy state, glucose and phospholipid metabolism.Key results:We found, by Western blotting and flow cytometry, that the levels of Bcr-Abl tyrosine kinase and multi-drug resistance p-glycoprotein were inconsistent among resistant clones. On the other hand, phospholipid metabolism and lactate production were highly predictive for cell response to imatinib. As previously reported, sensitive cells showed significantly decreased glycolytic activity (lactate) and phospholipid synthesis (phosphocholine) as well as increased phospholipid catabolism (glycerophosphocholine) after 24 h of 1 mu M imatinib treatment, which correlated with inhibition of cell proliferation and induction of apoptosis. In contrast to their sensitive counterparts, the K562-r, K562-R and LAMA84-r maintained increased phospholipid synthesis and glycolytic lactate production in the presence of 1 mu M (K562-r and LAMA84-r) and 5 mu M (K562-R) imatinib.Conclusions and implications:Specific metabolic markers for early detection of imatinib resistance, including increased glycolytic activity and phospholipid turnover, can be identified in resistant clones. Once validated in human isolated leukocytes, they may be used to monitor the responsiveness of CML patients to treatment.