氨磷汀对人巨核细胞白血病Dami细胞促分化作用的机制研究

Mechanism research of amifostine on inducing the differentiation of human megakaryoblastic leukemia Dami cell line

目的:探索氨磷汀对人巨核细胞白血病Dami细胞的促分化作用的机制。方法:采用氨磷汀1 mmol·L^(-1)刺激人巨核细胞白血病Dami细胞系,共诱导12 d,利用RT-PCR及Western blot方法检测在诱导的0、4 d、8 d、12 d时巨核细胞分化相关转录因子GATA-1、NF-E2的表达变化,并通过对Dami细胞进行核质分离以及利用免疫荧光的方法研究两种转录因子在核质中的分布。结果:RT-PCR及Western blot没有检测到巨核细胞分化相关转录因子GATA-1和NF-E2表达水平升高,但入核实验结果表明转录因子NF-E2、GATA-1入核活性增加。结论:氨磷汀诱导Dami细胞向成熟巨核细胞分化是通过增加转录因子NF-E2、GATA-1入核活性而发挥作用。

Objective:To explore the mechanism of amifostine on inducing the differentiation of human megakaryoblasticleukemia Dami cell line.Methods:The human megakaryoblastic leukemia Dami cell line were exposed to amifostine(1mmol·L-1)for12days.At different time points(0、4d、8d、12d),the expression of megakaryocytic differentiation-associated transcriptionfactors such as GATA-binding factor1(GATA-1)and nuclear factor erythroid2(NF-E2)were detected by RT-PCR and Western blot.Then in order to measure the distribution of the two transcription factors in nucleoplasm,nuclear and cytoplasmic were isolated,andimmunofluorescence was used.Results:There was no elevation on the mRNA or protein levels of megakaryocytic differentiationassociatedtranscription factors GATA-1and NF-E2by RT-PCR and Western blot,but nuclear import assay revealed an increasednuclear translocation activity of GATA-1and NF-E2.Conclusion:Amifostine induced the differentiation of Dami cells into maturemegakaryocytes via the mechanism involving increased nuclear translocation activity of the transcription factors GATA-1and NF-E2.