干扰素联合全反式维A酸逆转NB4-R1细胞的耐药机制研究

Effect of interferon combined with ATRA on differentiation of ATRA resistant NB4-R1 cells

目的:探讨干扰素(IFN)联合全反式维A酸(ATRA)逆转维甲酸耐药急性早幼粒细胞白血病(APL)的机制。方法:取指数生长期的NB4-R1细胞分为6组,分别加入ATRA、IFN-α、IFN-γ、IFN-α+ATRA、IFN-γ+ATRA(其中ATRA终浓度为2μmol/L,IFN-α和IFN-γ终浓度均为1 000IU/ml),以及不加药组的空白对照组,用MTT法检测细胞的增殖状况;蛋白免疫印迹试验测定各组NB4-R1细胞2hIRF-1蛋白表达,以及IFN-α+ATRA组、IFN-γ+ATRA组诱导NB4-R1细胞观察8hIRF-1蛋白表达的差异。结果:除ATRA组外,另外4个实验组均对NB4-R1细胞增殖有抑制作用,且联合用药的效果明显高于单独用药;5个实验组IRF-1蛋白均有表达,IFN-α+ATRA组、IFN-γ+ATRA组明显高于其他3组;IFN-α+ATRA组和IFN-γ+ATRA组比较,2h内前者IRF-1蛋白表达量逐渐下降,而后者则无明显变化;至用药8h后,IFN-α+ATRA组IRF-1蛋白表达基本消失,而IFN-γ+ATRA组的表达量仍较前无明显变化。结论:ATRA联合2种干扰素诱导NB4-R1细胞分化均有作用,但ATRA+IFN-γ效果更加显著;而IFN-α+ATRA组与IFN-γ+ATRA组比较,前者作用维持时间更长,可能与IRF-1蛋白持续表达有关。

Objective：To investigate the mechanism of interferon（IFN）combined with all-trans retinoic acid（ATRA）in reversing retinoic acid-resistant acute promyelocytic leukemia（APL）.Method：The experiment was divided into 6groups,respectively ATRA,IFN-α,IFN-γ,IFN-α＋ATRA and IFN-γ ＋ATRA（the final concentration of ATRA was 2μmol/L,IFN-αand IFN-γconcentration was 1 000IU/m）and the blank control group without any drugs,cell proliferation was tested with MTT assay,Western Blot was used to determine the IRF-1protein expression of NB4-R1 cells in 2hours,IRF-1protein expression of NB4-R1 cells in IFN-α＋ATRA group,IFN-γ＋ATRA group in 8hours.Result：Except ATRA group,the other 4experimental groups could inhibit the proliferation of NB4-R1 cells,and the combined treatment effect was significantly higher than that of single drug.The IRF-1protein expressed in 5experimental groups,and IFN-α＋ATRA group,IFN-γ＋ATRA group was significantly higher than those of the other 3groups.The expression of IRF-1protein in IFN-α＋ATRA group decreased gradually within 2hours,while IFN-γ＋ATRA group had no obvious change.Eight hours after the treatment,expression of IRF-1 protein in IFN-α＋ ATRA group disappeared,whereas IFN-γ＋ ATRA group still had no obvious change.Conclusion：ATRA combined with either kind of interferon plays a role in inducing NB4-R1 cell differentiation,but ATRA＋IFN-γhas more significant effects.The effect of IFN-α＋ATRA group has longer duration when compared with IFN-γ＋ATRA group,and the expression of IRF-1protein may be sustained.