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

摘要

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-l 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 staining and AnnexinV/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 μmol/L) inhibited the proliferation of NB4-R2 cells in both time- and concentration-dependent manners. The IC50 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 (jimol/L) induced 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 μmol/L) induces apoptosis on NB4-R2 cells, accompanied with reduction of MDR1 levels.%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 - 0.5 μmol/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 -5 μmol/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 -0.5 μmol/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0.064 μmol/L;0.05 -5 μmol/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( ＜0.5 μmol/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.