地西他滨对NB4及K562细胞增殖和凋亡的影响

摘要

本研究旨在观察去甲基化药物地西他滨(DAC)对NB4及K562细胞增殖和凋亡的影响.用台盼蓝染色法检测DAC对NB4及K562细胞的增殖抑制作用；流式细胞术检测不同浓度DAC作用后细胞周期的变化和CD11b的表达水平；瑞氏染色法观察药物作用后细胞形态变化；DNA梯形片段电泳分析细胞凋亡.结果表明,DAC显著抑制NB4及K562细胞的增殖(P＜0.05),且呈浓度和时间依赖性,DAC作用NB4及K562细胞72 h的半数抑制浓度(IC50)分别为0.113 μmol/L和0.138 μmol/L;不同浓度DAC作用72 h后,0.15 μmol/L DAC处理组两种细胞株G0/G1期细胞比例均显著增高,而S期细胞比例降低(P＜0.05)；两种细胞株的髓系分化抗原CD11b表达水平均升高(P＜0.05)；两种细胞株经不同浓度DAC处理48 h后,0.15μmoVL DAC处理组均出现典型的凋亡梯形DNA条带.结论:DAC抑制NB4及K562细胞增殖,诱导细胞分化,促进细胞凋亡；DAC对NB4细胞作用更明显.%This study was aimed to investgate the effects of decitabine (DAC) on proliferation and apoptosis of leukemia NB4 and K562 cells. The proliferation inhibition of DAC on NB4 and K562 cells was detected by Trypan blue staining. After treatment of DAC at different concentrations, the changes of cell cycle and CD11b expression was determined by flow cytometry. The cell morphological changes were observed by Wright's staining. The DNA ladder was used to detect cell apoptosis. The results indicated that DAC significantly inhibited the proliferation of NB4 and K562 cells in dose-and time-dependent manner. The median inhibitory concentration (IC50) of DAC-treated NB4 and K562 cells for 72 h was 0.113 μmol/L and 0.138 μmol/L, respectively. After treating these two cell lines with DAC at different concentration for 72 h, the cell ratio in G0/G1 phase significantly increased, while the cell ratio in S phase obviously decreased in 0. 15 μmol/L DAC group (P < 0. 05 ). The expression levels of myeloid differentiation antigen CDllb of both cell lines significantly increased in contrast to the control group (P < 0. 05 ), The cell morphology detected by Wright's staining displayed partial differentiation and apoptosis after treating NB4 and K562 cells with DAC for 48 h. Typical apoptotic DNA ladder was observed in 0.15 μmol/L DAC group at 48 h. It is concluded that DAC can inhibit NB4 and K562 cell proliferation, induce cell differentiation and apoptosis,but more obviously for NB4 cells.