新型维甲酸衍生物诱导NB4细胞分化的初步研究

目的:本研究探讨10种新型维甲酸衍生物对白血病细胞株NB4的抑制增殖和诱导分化活性。方法:维甲酸类衍生物作用于NB4细胞后,通过MTT法检测细胞的增殖。瑞氏染色法在倒置相差显微镜下观察加药处理前后细胞形态学变化。NBT还原实验法分析细胞的分化指标。FCM检测分析细胞周期和细胞表面分化抗原变化。结果:维甲酸衍生物作用3 d后,抑制细胞增殖作用呈剂量依赖效应。10种维甲酸衍生物(10-5mol/L)的诱导分化活性表现在油镜下观察NB4细胞向粒系分化成熟的改变,NBT阳性细胞率增加,细胞表面分化抗原CD11b表达量增加,CD13表达则减少。通过对细胞周期的分析,发现G0/G1期细胞表达量增加,呈G1期阻滞。结论:维甲酸衍生物2a-03,4a-02和5a-02显示有较强的诱导NB4细胞分化的活性。

Preliminary study of NB4 cell differentiation induced by retinoic acid derivatives

AIM: To investigate the proliferation inhibition and differentiation induction activities of 10 retinoic acid derivatives in NB4 cells,a human promyelocytic leukemia cell line.METHODS: Cell proliferation was assessed by MTT assay before and after the treatment with retinoic acid derivatives in vitro.Morphologic changes were observed after Wright's staining using inverted phase contrast microscope.Cell differentiation indexs were analyzed by nitroblue tetrazolium(NBT) reduction test.Cell cycle and the expression of the maturation specific cell surface markers CD11b,CD13 were determined by Flow Cytometry(FCM) analysis.RESULTS: The growth of NB4 cells was inhibited in a does-dependent manner after the treatment with retinoic acid derivatives for 3 days.The changes of differentiation and maturation of NB4 cells induced by 10 retinoic acid derivatives at the concentration of 10-5 mol/L were observed through immersion objective.NBT reduction test indicated that the retinoic acid derivatives could induce differentiation of NB4 cells and increase the positive cell ratio.The expression level of the maturation specific cell surface marker CD11b was increased while the expression level of CD13 was decreased.The proportion of cells in G0/G1 phase was increased.Cell cycle progression was blocked in the G1 phase.CONCLUSION: The retinoic acid derivatives retinoid 2a-03,4a-02 and 5a-02 have powerful activity in inducing NB4 cells differentiation.