BET bromodomain蛋白在人胚胎干细胞造血分化中的作用研究

Role of BET Bromodomain in Hematopoietic Differentiation from hESCs

目的:探讨BET bromodomain蛋白在人胚胎干细胞造血分化中的作用。方法:应用人胚胎干细胞单层造血分化体系,通过免疫荧光技术、流式细胞术及实时定量PCR技术,研究BET bromodomain蛋白抑制剂I-BET151对造血分化的影响,并在不同分化阶段添加I-BET 151探讨其在造血分化过程中的作用阶段。结果:I-BET151能够显著抑制CD43^+造血干/祖细胞的产生,并且在APLNR^+侧板中胚层生成、CD34^+CD31^+生血内皮产生及内皮细胞生血转化3个阶段添加I-BET 151对CD43^+造血干/祖细胞的产生均有显著抑制作用。结论:人胚胎干细胞造血分化过程中添加BET bromodomain蛋白抑制剂I-BET 151,能够抑制CD43^+造血干/祖细胞的产生,并进一步证实BET

Objective： To explore the role of bromodomain and extra terminal（ BET） bromodomain in hematopoietic differentiation from human enbryonic stem cells（ hESC）. Methods： The effect of BET hematopoietic inhibitor I-BET151 on hematopoietic differentiation from hESC was detected by using a monolayer hematopoietic defferentiation model,immunofluorescence,flow cytometry and real-time PCR; moreover the role of I-BET151 in process of hematopoietic differentiation was explored by adding I-BET151 in different differentiation stages. Results： The analysis results of immunofluorescence,flow cytometry and real-time PCR showed that I-BET 151 significantly inhibited the generation of CD43 positive hematopoietic stem and progenitor cells（ HSPCs）. It was found that the addition of I-BET151 in different stages,including APLNR ＋ lateral plate mesoderm production,CD34 ＋ CD31 ＋ hemogenic endothelium（ HEP） generation and endothelial-to-hematopoietic transition,significantly suppressed the generation of CD43 positive hematopoietic progenitor cells. Conclusion： I-BET 151 inhibites hematopoietic differentiation from hESCs at several stages,suggesting that the BET bromodomain plays important roles in multiple stages of hematopoietic differentiation from hESCs.