硫氧还蛋白在急性髓系白血病中的表达及其抑制剂体外对急性髓系白血病细胞促凋亡作用的初步研究  

Preliminary study on thioredoxin expression in AML cells and apoptosis of AML cells induced by diamide in vitro

作　　者：许莲蓉[1] 冯江芳[1] 牛勃[1] 陈显久[1] 常冰梅[1] 胥显民[1] 

机构地区：[1]山西医科大学第二医院血液科,太原030001

出　　处：《白血病．淋巴瘤》2011年第12期719-722,共4页Journal of Leukemia & Lymphoma

摘　　要：目的探讨急性髓系白血病（AML）细胞中硫氧还蛋白（Trx）的表达及其与AML患者临床特点及预后的相关性，观察Trx抑制剂二酰胺体外诱导白血病细胞凋亡的作用，为临床治疗AML提供一定的理论依据。方法选择20例AML患者和10名健康人，抽取其骨髓，分离单个核细胞。用Westernblot法分析单个核细胞中Trx的表达，应用四甲基偶氮唑蓝比色（MTT）法检测不同浓度二酰胺对单个核细胞增殖的影响，应用形态学、琼脂糖凝胶电泳观察二酰胺对细胞凋亡的诱导作用。结果AML患者Trx的表达率为75％（15／20），在健康人中不表达。Trx与患者的白细胞计数显著相关（X2=9．375，P〈0．05）。MTT法检测显示二酰胺对细胞具有细胞毒作用，且呈时间和剂量依赖性，24、48和72h的‰值分别为98．26、47．53、8．34mg／ml。形态学观察二酰胺作用于AML细胞后可见凋亡小体，琼脂糖凝胶电泳观察可见典型的DNA梯形条带出现。结论AML细胞Trx含量明显高于健康人，二酰胺干预后AML细胞生长受抑甚至发生凋亡，其可能成为AML治疗的新药。Objective To explore the expression level of thioredoxin （Trx） in acute myeloid leukemia （AML） cells, and its association with clinical features and prognosis, as well as the effects of diamide, an inhibitor of Trx, in inducing AML cells apoptosis. Methods Expression of Trx on AML cells and fresh bone marrow cells from healthy adults were analyzed by Western-blotting. The inhibition of proliferation was measured by MTY assay. The anti-leukemia effect of diamide was observed by morphology and agarose gel electrophoresis. Results 75 % （15/20） of AML patients expressed Trx, while no expression was observed in control group. Higher expression level of Trx was associated with higher WBC counts （X2 = 9.375, P 〈 0.05）, which suggested that overexpression was associated with leukemogenesis. The inhibition of diamide on AML cells showed time and dose dependent by MTr assay. The IC5o values of diamide at 24 h, 48 h and 72 h were 98.26 mg/ml, 47.53 mg/ml and 8.34 mg/ml, respectively. After AML cells were treated with diamide, the apoptotic body appeared by morphology, and the typical DNA ＂ladder＂ bands were confirmed by agarose gel electrophoresis. Conclusion Trx expression level in AML ceils is significantly higher than that of control group. Diamide inhibites the proliferation of AML cells by inducing apoptosis, which might be a potential agent for AML.

关 键 词：白血病 实验性 白血病 急性 硫氧还蛋白 二酰胺 细胞凋亡 

分 类 号：R733.7[医药卫生—肿瘤]