丹参酮与三氧化二砷协同诱导急性早幼粒细胞白血病细胞株凋亡的体外研究

Study of combination of tanshinone with arsenic trioxide induced apoptosis of acute promyelocytic leukemia cells in vitro

目的为评估丹参酮ⅡA(TanshinoneⅡA,TanⅡA)与三氧化二砷(As2O3)联合应用的效应,观察2药的相互作用和相互影响,为其临床应用提供依据。方法以不同浓度的As2O3、TanⅡA单独及联合应用作用于急性早幼粒细胞白血病细胞(NB4)作为实验组,通过观察细胞形态学改变,以MTT法测定细胞存活率,并采用流式细胞术(FCM)检测细胞周期分布,分析不同浓度的As2O3、TanⅡA单独及联合应用对NB4细胞的增殖、凋亡的影响。结果 TanⅡA、As2O3均能抑制NB4细胞生长,以1.0μmol/L As2O3作用更显著,二者联合应用后的增殖抑制作用表现为协同效应;经TanⅡA组处理后的NB4细胞,其形态改变更多显示一定程度的成熟粒细胞的特征,As2 O3组则更多表现为细胞凋亡的形态学改变,并经电镜检查证实,两药联合应用可加强NB4细胞的凋亡形态改变;处理后的NB4细胞G0/G1期比例增高,S期细胞比例降低,细胞增殖指数(PI)下降,两药联合应用后具有协同效应。结论①TanⅡA、As2O3对NB4细胞均有增殖抑制作用,两者的增殖抑制作用具有协同效应,并与剂量相关;②TanⅡA与As2O3联合作用于NB4细胞具有诱导凋亡作用,两药的诱导凋亡作用具有协同效应;③小剂量的As2 O3联合TanⅡA即可对NB4细胞产生促进凋亡的效应,故临床可以选择小剂量的As2 O3,以减轻砷剂的毒副作用。

Objective To assess effects of TanⅡA combined the As2O3,and to observe interaction of two drug and to provide a theoretical basis for clinical application.Methods According to different concentrations of As2O3,TanⅡA used alone and combination for the United NB4 cell line as the experimental group.The effects were analyzed in different concentrations of As2O3,TanⅡA used alone and combination of NB4 cell proliferation and apoptosis by observing the cell morphological changes,MTT cell proliferation survival experiments,using flow cytometry（FCM）detection cell cycle distribution.Results TanⅡA,As2O3 could inhibit the growth of NB4 cells to 1.0μmol/L As2O3,and use of the combination showed synergistic growth inhibition effect;In TanⅡA group of NB4 cells after treatment,the morphological changes showed a certain degree of maturation of myeloid features,and As2O3 group showed more morphological changes of apoptosis,and by electron microscopy confirmed that the two drugs in combination could enhance the apoptosis of NB4 cells to change shape;NB4 cells after treatment increased the proportion of G0/G1 phase,and S phase cells reduced the proportion of cell proliferation index（PI）decreased after the two drugs in combination with a synergistic effect.Conclusion There is inhibition for proliferation of NB4 cells by TanⅡA and As2O3;As2O3 combined TanⅡA has synergistic effect with the inhibition and dose-dependent and TanⅡA combined As2O3 can induce apoptosis in NB4 cells;Small doses of As2O3 combined TanⅡA can promote apoptosis of effect for NB4 cells.