Jilin University China-Japan Union Hospital, Changchun, 130033 China.
Jilin University China-Japan Union Hospital, Changchun, 130033 China ; Tumor Hospital of Jilin Province, Changchun, 130021 China.
Proteome Sci. 2015 Jan 22;13(1):1. doi: 10.1186/s12953-014-0057-y. eCollection 2015.
Chemoresistance remains a significant challenge in chronic myelogenous leukemia (CML) management, which is one of the most critical prognostic factors. Elucidation the molecular mechanisms underlying the resistance to chemoresistance may lead to better clinical outcomes.
In order to identify potential protein targets involved in the drug-resistant phenotype of leukemia, especially the chronic myelogenous leukemia (CML), we used a high-resolution "ultra-zoom" 2DE-based proteomics approach to characterize global protein expression patterns in doxorubicin-resistant myelogenous leukemia cells compared with parental control cells. Ultra-high resolution of 2DE was achieved by using a series of slightly overlapping narrow-range IPG strips during isoelectric focusing (IEF) separation. A total number of 44 proteins with altered abundances were detected and identified by MALDI-TOF or LC-MS/MS. Among these proteins, enolase, aldolase, HSP70 and sorcin were up-regulated in doxorubicin-resistant myelogenous leukemia cell line, whereas HSP27 was down-regulated. Some of the results have been validated by Western blotting. Both enolase and aldolase were first reported to be involved in chemoresistance, suggesting that process of glycolysis in doxorubicin-resistant myelogenous leukemia cells was accelerated to some extent to provide more energy to survive chemical stress. Possible roles of most of the identified proteins in development of chemoresistance in myelogenous leukemia cells were fully discussed. The results presented here could provide clues to further study for elucidating the mechanisms underlying drug resistance in leukemia.
As a whole, under the chemical stress, the doxorubicin-resistant myelogenous leukemia cells may employ various protective strategies to survive. These include: (i) pumping the cytotoxic drug out of the cells by P-glycoprotein, (ii) increased storage of fermentable fuel, (iii) sophisticated cellular protection by molecular chaperones, (iv) improved handling of intracellular calcium, (v) increased glucose utilization via increased rates of glycolysis. In the present study, proteomic analysis of leukemia cells and their drug resistant variants revealed multiple alterations in protein expression. Our results indicate that the development of drug resistance in doxorubicin-resistant myelogenous leukemia cells is a complex phenomenon undergoing several mechanisms.
化学耐药性仍然是慢性髓细胞白血病（CML）管理中的一个重大挑战，这是最关键的预后因素之一。阐明导致耐药性的分子机制可能会带来更好的临床结果。
为了鉴定参与白血病耐药表型的潜在蛋白靶标，特别是慢性髓细胞白血病（CML），我们使用高分辨率的“超聚焦”二维电泳（2-DE）基于蛋白质组学方法来比较多柔比星耐药髓系白血病细胞与亲本对照细胞之间的全局蛋白表达模式。在等电聚焦（IEF）分离过程中使用一系列略微重叠的窄范围 IPG 条带实现了 2-DE 的超高分辨率。通过 MALDI-TOF 或 LC-MS/MS 检测和鉴定了 44 个丰度改变的蛋白。其中，烯醇酶、醛缩酶、HSP70 和 sorcin 在多柔比星耐药髓系白血病细胞系中上调，而 HSP27 下调。其中一些结果已通过 Western blot 验证。烯醇酶和醛缩酶均首次报道参与耐药性，表明多柔比星耐药髓系白血病细胞的糖酵解过程在某种程度上加速，以提供更多的能量来应对化学应激。还充分讨论了鉴定的大多数蛋白在髓系白血病细胞耐药性发展中的可能作用。这里呈现的结果可以为进一步阐明白血病耐药机制的研究提供线索。
总的来说，在化学应激下，多柔比星耐药髓系白血病细胞可能采用各种保护策略来存活。这些策略包括：（i）通过 P-糖蛋白将细胞毒性药物泵出细胞，（ii）增加可发酵燃料的储存，（iii）通过分子伴侣进行复杂的细胞保护，（iv）改善细胞内钙的处理，（v）通过增加糖酵解速率增加葡萄糖利用。在本研究中，白血病细胞及其耐药变体的蛋白质组学分析揭示了蛋白表达的多种改变。我们的结果表明，多柔比星耐药髓系白血病细胞的耐药性发展是一个经历多种机制的复杂现象。
