白藓碱单用及联合全反式维甲酸对急性髓性白血病细胞的分化治疗作用

Single and Combined Effects of Dictamnine with All-trans Retinoic Acid(ATRA) on Differentiation of Acute Myeloid Leukemia(AML) Cells

关键词:  白藓碱  全反式维甲酸  急性髓性白血病  分化  转录因子

Key words:  dictamnine  all-trans retinoic acid(ATRA)  acute myeloid leukemia(AML)  differentiation  transcription factors

摘要:

目的 探索白藓碱单用及联合全反式维甲酸(ATRA)对急性髓性白血病细胞(AML)的分化治疗作用。方法 不同浓度的白藓碱单独或联合ATRA作用于3株不同AML亚型的细胞株(NB4、U937和HL60)，利用细胞计数考察细胞增殖抑制情况；通过PI染色结合流式细胞术检测细胞凋亡；检测细胞表面抗原CD11b和细胞核形态考察细胞分化；RT-PCR和Western blotting法观察细胞分化相关转录因子CEBPβ和PU.1的mRNA和蛋白水平。结果 白藓碱单用及联合ATRA均能明显诱导AML细胞发生分化，表现为抑制细胞增殖、增加细胞表面CD11b的表达，使细胞核形态出现马蹄形甚至分叶形；白藓碱单用及联合ATRA均能在mRNA水平和蛋白水平上增加分化相关转录因子CEBPβ和PU.1的表达。结论 白藓碱能够单独或协同ATRA诱导不同AML亚型细胞发生分化，并且其诱导分化作用可能是通过调控分化相关转录因子CEBP?和PU.1引起的。

Abstract:

OBJECTIVE To explore the single and combined effects of dictamnine with all-trans retinoic acid (ATRA) on acute myeloid leukemia(AML) cells. METHODS Cell proliferation assay was performed by manual cell counting in Burker chambers; apoptosis was detected by flow cytometry with PI staining; CD11b expression level and cell morphologic changes were used to determine AML cell differentiation. The expression levels of transcription factors CEBPβ and PU.1 were analyzed by western blotting and real-time PCR. RESULTS Both dictamnine single treatment and its combination treatment with ATRA could inhibit AML cell proliferation and induce CD11b expression and morphologic changes, while no obvious apoptosis was observed. Comparing with ATRA or dictamnine single treatment, the mRNA and protein levels of CEBPβ and PU.1 were up-regulated in dictamnine and ATRA cotreated AML cells. Meanwhile, dictamnine alone also had the same effect to induce the expression of CEBPβ and PU.1. CONCLUSION Dictamnine alone can induce AML cell differentiation and also contribute to the ATRA-induced differentiation of AML cells, and the activation of CEBPβ and PU.1 are involved in this process.