Eos对红系分化和巨核系分化的调控作用及机制研究

Eos is a member of the zinc ?nger transcription factor Ikaros family and play a pivotal role as a repressor in regulation of several important genes. Our result showed that Enforced expression of Eos can inhibit Hemin-induced K562 cells erythroid differentiation and Epo-induced CD34+ HPCs erythroid differentiation. Recently, we found the obvious decrease of Eos expression by Western during PMA-induced K562 cells megakaryotic differentiation, so we speculate that Eos will inhibit megakaryotic differentiation.And there are unclear the mechanism how Eos affect erythroid and megakaryotic differentiation. But the expression of the important transcription factor GATA-1 was changed when Eos was enforced expression in K562 cells.So Eos may be involved in erythroid and megakaryotic differentiation through interacting with GATA-1 protein. By using the methods of MGG staining, flow cytometry, co-Immunoprecipitation, RNAi silence etc.,we will firstly discover that the function of Eos in megakaryotic differentiation, and confirm the important functional interaction between Eos and GATA-1. Taken together, our observations will demonstrate that Eos play an important role in in erythroid and megakaryotic differentiation. Our results will supply the significant clues for the detailed mechanisms and regulation network of erythroid differentiation and megakaryotic differentiation, all of these will possibly provide potential targets for studying and curing the diseases related to hematopoietic differentiation.

Eos也被称作IKZF4，是锌指蛋白转录因子ikaros家族中的一员，它作为负调节因子参与了多种重要基因表达调控。我们已证实Eos可以抑制K562细胞和CD34+造血干/祖细胞红系分化，最近发现Eos在K562细胞巨核系分化过程中表达下调，因此推测Eos对巨核系分化可能也有抑制作用。Eos对红系和巨核系分化的作用机制如何？目前尚不清楚。但Eos不直接影响红系和巨核系分化关键转录因子GATA-1基因表达，那么Eos可能通过与GATA-1相互作用参与到红系和巨核系分化中。本项目拟采用MGG染色、流式细胞仪检测、免疫共沉淀和RNAi等技术，将首次证实Eos对巨核系分化具有重要作用，并拟揭示Eos与GATA-1之间存在相互作用。总之，本研究将深入探讨Eos在红系和巨核系分化中的重要功能，为复杂的造血分化调控网络找到新的线索，并对造血分化相关疾病研究和治疗提供潜在的靶点。

Eos也被称作IKZF4（ikaros family zinc finger 4）是锌指蛋白转录因子ikaros家族中的一员，它作为一个负调节因子参与到多种基因的表达调控。我们的研究发现Eos可以抑制Hemin诱导的K562细胞的红系分化和Epo诱导的CD34+造血干/祖细胞的红系分化。同时，Eos也可以抑制PMA诱导的K562细胞的巨核细胞分化和Tpo诱导的CD34+造血干/祖细胞的巨核细胞分化。我们拟采用MGG染色、流式细胞仪检测、免疫共沉淀、RNAi、Real-time PCR和Western blot等技术系统地研究Eos对红系分化和巨核系分化的调节作用及其机制。本研究首次揭示Eos在巨核系分化中的重要作用，并证实了Eos与造血转录因子GATA-1之间存在重要相互作用。总之，本研究结果将为复杂的红系和巨核系造血分化调控网络找到新的线索，并对造血分化相关疾病的研究和治疗提供潜在靶点。