Role of GSTP1-1 in mediating the effect of As2O3 in the acute promyelocytic leukemia cell line NB4
Role of GSTP1-1 in mediating the effect of As2O3 in the acute promyelocytic leukemia cell line NB4
复制标题
DOI：
10.1007/s00277-006-0139-8
复制
发表时间：
2006-10-01
影响因子：
3.5
通讯作者：
Lo Coco, Francesco
中科院分区：
文献类型：
--
作者：
Bernardini, Sergio;Nuccetelli, Marzia;Lo Coco, Francesco
三氧化二砷（As_2O_3）是治疗急性早幼粒细胞白血病（APL）的一种高效药物，而其他造血系统肿瘤对该药物的反应性较低，并且对As_2O_3耐药的机制知之甚少。为了更好地了解GSH相关途径在As 2 O3敏感性中的复杂网络，我们研究了GSH和GSH相关酶在对As 2 O3敏感的APL细胞系（NB 4）和耐药亚克隆（AsR）中的作用。细胞增殖，活力和凋亡进行了研究，在NB 4细胞治疗前和治疗后，与1 μ M As 2 O3和AsR细胞。在这些实验细胞模型中，通过免疫印迹分析GSTP 1 -1、JNK 1和JNK 2蛋白，并进行JNK 1的激酶测定。GSH水平以及酶谷胱甘肽过氧化物酶、谷胱甘肽转移酶、γ-谷氨酰半胱氨酸转氨酶和超氧化物歧化酶的活性被测量。As 2 O3处理的NB 4细胞表现出高水平的氧化应激和GSH水平的增加。GSTP 1 -1聚合和JNK 1活化在24小时后可检测到，随后在72小时开始凋亡率增加。既没有GSTP 1 -1聚合，也没有JNK激活的AsR细胞，表现出非常低的凋亡率。我们的研究结果表明，APL的敏感性As 2 O3可能，至少部分，介导的协会和从GSTP 1 -1的JNK解离之间的平衡，这取决于细胞的氧化还原状态。进一步的研究是必要的，以找到一种方法来干扰这种平衡，每当它可能代表一种耐药机制。
Arsenic trioxide (As2O3) is a highly effective agent in the treatment of acute promyelocytic leukemia (APL), whereas other hematopoietic tumors are less responsive to this agent and mechanisms underlying As2O3,-resistance are poorly understood. To better understand the complex network of GSH-related pathways in As2O3 sensitivity, we investigated the role of GSH and GSH-relevant enzymes in an APL cell line sensitive to As2O3 (NB4) and in a resistant subclone (AsR). Cell proliferation, viability, and apoptosis were investigated in NB4 cells before and after treatment with 1 mu M As2O3 and in AsR cells. In these experimental cell models, GSTP1-1, JNK1 and JNK2 proteins were analyzed by immunoblotting, and a kinase assay for JNK1 was performed. GSH levels as well as the activities of the enzymes glutathione peroxidase, glutathione transferase, gamma-Glutamylcysteynilsinthetase and superoxide dismutase were measured. NB4 cells treated with As2O3 showed a high level of oxidative stress and an increase of GSH levels. GSTP1-1 polymerization and JNK1 activation were detectable after 24 h and were followed by an increase of the apoptotic rate starting at 72 h. Neither GSTP1-1 polymerization nor JNK activation was found in AsR cells that showed a very low apoptotic rate. Our results suggest that APL sensitivity to As2O3 might be, at least in part, mediated by the balance between association and dissociation of JNK from GSTP1-1, depending on the redox status of the cell. Further investigation is warranted to find a way to interfere with this balance, whenever it might represent a mechanism of drug resistance.