Characterization of cytarabine-resistant leukemic cell lines established from five different blood cell lineages using gene expression and proteomic analyses
Characterization of cytarabine-resistant leukemic cell lines established from five different blood cell lineages using gene expression and proteomic analyses
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DOI：
10.3892/ijo.2011.933
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发表时间：
2011-04-01
影响因子：
5.2
通讯作者：
Ueda, Takanori
中科院分区：
文献类型：
--
作者：
Negoro, Eiju;Yamauchi, Takahiro;Ueda, Takanori
阿糖胞苷(Ara-C)是治疗急性髓系白血病的关键药物。由于细胞内阿糖胞苷三磷酸(Ara-CTP)是阿糖胞苷-C的活性代谢物，因此减少阿糖胞苷-CTP含量的因素被认为是导致耐药性的因素。然而，这些因素并不能完全解释对阿糖胞苷产生抗药性的原因。本研究旨在寻找新的候选Ara-C抗性因子，包括那些与Ara-CTP生产无关的因子。为此，我们从不同血细胞系白血病细胞系(HL-60、K562、CEM、THP1和U937)中新建立了5个抗Ara-C的白血病克隆。抗性亚无性系的Ara-C抗性是亲本的5-58倍。除抗Ara-C的CEM细胞外，所有抗Ara-C的亚克隆都表现出Ara-CTP相关因子的改变，如Ara-C膜转运能力、脱氧胞苷酶活性或胞质核苷酸酶11活性。为了确定新的候选因子，我们使用了两种综合方法：DNA微阵列和蛋白质组分析。DNA芯片分析发现，与亲本细胞相比，5个Ara-C抗性株系中有8个基因(C19orf2、HSPA8、LGALS1、POU4F3、PSAP、AKT1、MBC2和CACNA2D3)发生了改变。蛋白质组和DNA芯片分析进一步检测到，与其亲本系相比，抗Ara-C的CEM亚克隆中stathmin1的蛋白水平降低。因此，本研究结果提示多种新的机制参与了白血病细胞对阿糖胞苷耐药的调节。其中一些分子在抗药性中的作用尚不清楚。
Cytarabine (ara-C) is the key drug for treatment of acute myeloid leukemia. Since intracellular cytarabine triphosphate (ara-CTP) is an active metabolite of ara-C, factors that reduce the amount of ara-CTP are known to induce drug resistance. However, these factors do not fully explain the development of resistance to ara-C. The present study was conducted to search for new candidate ara-C resistance factors, including those that are unrelated to ara-CTP production. For this purpose, we newly established five ara-C-resistant leukemic clones from different blood cell lineage leukemic cell lines (HL-60, K562, CEM, THP1 and U937). The resistant subclones were 5-58-fold more ara-C-resistant than their parental counterparts. All of the ara-C-resistant subclones, except for ara-C-resistant CEM cells, displayed alteration of ara-CTP-related factors such as ara-C membrane transport capacity, deoxycytidine kinase activity or cytosolic nucleotidase 11 activity. To identify new candidate factors, we used two comprehensive approaches: DNA microarray and proteome analyses. The DNA microarray analysis revealed eight genes (C19orf2, HSPA8, LGALS1, POU4F3, PSAP, AKT1, MBC2 and CACNA2D3) that were altered in all five ara-C-resistant lines compared to parental cells. Both proteome and DNA microarray analyses further detected a reduced protein level of stathmin 1 in the ara-C-resistant CEM subclone compared to its parental line. Thus, the present findings suggested the involvement of novel multiple mechanisms in mediating the ara-C resistance of leukemic cells. The role of some of these molecules in resistance is still unclear.