四硫化四砷诱导维甲酸耐药的急性早幼粒白血病NB4-R1细胞凋亡及其分子机制  

Apoptosis of Retinoic Acid-Resistant APL NB4-R1 Cells Induced by As_4S_4 and Its Molecular Mechanism

作　　者：王嫄[1] 张梅[1] 贺鹏程[1] 刘雁峰[1] 齐珺[1] 程小岩[1] 朱华超[1] 

机构地区：[1]西安交通大学医学院第一附属医院血液内科,陕西西安710061

出　　处：《中国实验血液学杂志》2015年第1期83-87,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金项目资助(81000218)

摘　　要：目的:探讨四硫化四砷(As4S4)对维甲酸耐药的NB4-R1细胞诱导凋亡的作用及其机制。方法:As4S4作用于NB4-R1细胞0、24、48 h后,收集细胞,按照不同的处理时间分为对照组和实验组。应用流式细胞仪检测细胞周期及凋亡,DNA琼脂糖凝胶电泳检测DNA降解断裂条带,蛋白凝胶电泳Western blot方法检测药物作用NB4-R1细胞后BCL-2、BAX和Caspase-3蛋白的表达。结果:25μmol/L As4S4作用0、24和48 h后,NB4-R1细胞早期凋亡率由0%升至24.49%和47.41%,晚期凋亡率由0.08%升至14.72%和20.70%,细胞凋亡呈现时间依赖性。应用As4S4处理24 h后可以观察到凋亡细胞DNA降解形成的明显的梯形条带。细胞周期分析发现,25μmol/L As4S4能使NB4-R1细胞阻滞于S期,药物作用24 h及48 h后,S期细胞由31.85%升高至42.53%及55.12%,G0/G1期细胞由57.30%降至了37.56%及28.51%。Western blot检测发现,As4S4作用NB4-R1细胞48 h后,BAX表达上调,BCL-2表达下调,Caspase-3出现活化条带。结论:As4S4能有效的诱导NB4-R1细胞凋亡,并通过影响BCL-2、BAX、Caspase-3蛋白表达,触发线粒体凋亡途径诱导细胞凋亡。Objective： ： This study was to investigate the apoptosis-inducing effect of As4S4 on the retinoic acid- resistant acute promyelocytic leukemia（APL） NB4-R1 cells and its potential mechanisms. Methods： The leukemia cell line NB4-R1 was cultured in vitro and divided into control group and treatment group. The apoptosis rate and cell cycle were detected by flow cytometry. The apoptotic DNA fragments were analyzed by agarose gel electrophoresis. The changes of BCL-2, BAX and Caspase-3 were determined by Western blot. Results： After NB4-R1 cells were treated with As4S4 （25 μmol/L） for 0 h, 24 h, 48 h, the percentage of early apoptotic cells was obviously raised from 0% to 24.49% and 47.41%, the percentage of late apoptotic cells were elevated from 0.08% to 14.72% and 20.70%. Compared with control group, the DNA degradation revealed a characteristic DNA ladder during agarose gel electrophoresis after treatment for 24 h. The drug significantly induced an accumulation of the S phase cell population from 31.85% of the untreated cells to 42.53% and 55.12% treated with the different time whereas the NB4-R1 cells in G0/G1 phase decreased from 57.30% to 37.56% and 28.51%. As4S4 could decrease the expression of BCL-2 and increase the level of BAX. Pro-caspase-3 could be cleaved into small active fragments under the apoptotic stimulation. Conclusion：As4S4 can efficiently induce NB4-R1 cell apoptosis, which may be related with the down-regulation of BCL-2 and the up-regulation of BAX, as well as the activation of Caspase-3.

关 键 词：急性早幼粒细胞白血病 NB4-R1细胞 AS4S4 耐药 细胞凋亡 

分 类 号：R733.71[医药卫生—肿瘤] R979.1[医药卫生—临床医学]