地西他滨对全反式维甲酸耐药细胞株NB4-R2的增殖抑制及诱导凋亡作用研究  

Decitabine Inhibits Cell Proliferation and Induces Apoptosis of All-trans Retinoid Acid-resistant Acute Promyelocytic Leukemia NB4-R2 Cell Line

作　　者：熊暮珺[1,2] 肖若芝[1,2] 陈琰[1] 陈家杰[2] 龙梓洁[1,2] 吴星[2] 林东军[1,2] 

机构地区：[1]中山大学附属第三医院血液科 [2]中山大学血液病研究所,广东广州510630

出　　处：《中国实验血液学杂志》2012年第1期48-52,共5页Journal of Experimental Hematology

摘　　要：本研究探讨地西他滨(DAC)对全反式维甲酸耐药细胞株NB4-R2的增殖抑制及诱导凋亡作用。应用细胞增殖及毒性检测法(新型四唑单钠盐法)观察0.01-0.5μmol/L DAC作用于NB4-R2细胞24、48及72 h后的细胞增殖活力变化;PI单染及AnnexinⅤ-FITC/PI双染流式细胞术观察不同浓度DAC(0.05-5μmol/L)对NB4-R2处理48 h后的细胞凋亡率;RT-PCR法检测编码P糖蛋白(P-gp)的多药耐药基因1(MDR1)转录水平的变化。结果表明,0.01-0.5μmol/L DAC可抑制NB4-R2细胞增殖,并呈现明显的量-效与时-效关系;作用48及72 h后的IC50分别为0.089和0.064μmol/L;0.05-5μmol/L DAC以浓度依赖的方式诱导NB4-R2细胞凋亡,下调MDR1基因表达。结论:低浓度DAC(<0.5μmol/L)对NB4-R2细胞有增殖抑制作用,较高浓度DAC(1、5μmol/L)可以诱导NB4-R2细胞凋亡,同时使MDR1表达下调。The aim of this study was to investigate the proliferation-inhibitory and inducing apoptotic effects of decit- abine （DAC） on acute promyelocytic leukemia NB4-R2 cells. Cell inhibitory rate was determined by cell proliferation and cytotoxicity assay（ WST-1 assay）after NB4-R2 cells were treated with 0.01 -0.5 μmol/L DAC for 24,48 and 72 h. Apoptosis of NB4-R2 cells treated with 0.05 - 5 μmol/L DAC for 48 h was detected by flow cytometry with PI stai- ning and Annexin V/PI staining. Reverse transcription-PCR （RT-PCR） was used to determine the mRNA expression level of MDR1 gene encoding P-glycoprotein （P-gp）. The results indicated that DAC （0.01 -0.5 μ mol/L） inhibited the proliferation of NB4-R2 cells in both time- and concentration-dependent manners. The ICs0 of DAC on the viability of NB4-R2 cells after treatment for 48 and 72 h were 0.089 and 0.064 μmol/L respectively. DAC （0.05 - 5 μmol/L） in- duced NB4-R2 cell apoptosis in dose-dependent manner with down-regulation of MDR1 gene expression. It is concluded that a low concentration of DAC （ 〈0.5 μmol/L） inhibites cell proliferation, while higher concentration of DAC （1 or 5 ixrnol/L） induces apoptosis on NB4-R2 cells, accompanied with reduction of MDR1 levels.

关 键 词：地西他滨 NB4-R2细胞 细胞凋亡 多药耐药基因 

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