AML1a在小鼠造血细胞增殖分化异常中的作用  

Role of AML1a in Abnormal Proliferation and Differentiation of Murine Hematopoietic Cells

作　　者：许发美 邢海燕[1] 田征[1] 唐克晶[1] 饶青[1] 王敏[1] 王建祥[1] 

机构地区：[1]中国医学科学院,北京协和医学院,血液学研究所血液病医院,实验血液学国家重点实验室,天津300020

出　　处：《中国实验血液学杂志》2011年第6期1477-1481,共5页Journal of Experimental Hematology

基　　金：天津市应用基础研究计划项目,编号10JCZDJC19600

摘　　要：本研究旨在探讨AML1a在小鼠造血细胞增殖分化异常中的作用及其作用机制。将构建的pMSCV-FLAG-AML1a-IRES-YFP和pMSCV-IRES-YFP分别与辅助质粒pV Pack-Eco(含Env)、pV Pack-GP(含gal-pol)组合,通过磷酸钙沉淀法转染293T细胞,制备逆转录病毒。通过逆转录病毒将AML1a及YFP转导至C57 BL/6J雄性小鼠的骨髓单个核细胞(BMMNC),接种于M3434甲基纤维素完全培养液进行集落形成实验,并置于含mSCF、mIL-3、mIL-6的M5300液体培养液中进行长期培养,观察BMMNC形态变化。结果显示,转导了AML1a的BMMNC集落形成能力增强,集落数量和体积均明显大于对照组,集落以CFU-E和CFU-GEMM为主。长期培养实验中,转导AML1a组细胞形态显示分化阻滞,而对照组细胞则处于更成熟的阶段。结论:AML1a增加了造血干/祖细胞的增殖能力,并将小鼠造血干/祖细胞阻滞于较早的发育阶段。This study was aimed to explore the role and mechanism of AML1a in abnormal hematopoiesis in mice.Plasmids pMSCV-FLAG-AML1a-IRES-YFP and pMSCV-IRES-YFP together with envelope-encoding plasmid pECO and packaging plasmid pGP were respectively transfected into 293T cells by using a method of calcium phosphate precipitation to produce retrovirus.Bone marrow mononuclear cells（BMMNC） from male C57BL/6J mice were transfected with the retroviral vector MSCV expressing FLAG-AML1a fusion protein and yellow fluorescent protein（YFP）.The cells were cultured in M3434 semi-solid medium for colony formation assay and in M5300 fluid medium containing murine IL-3（mIL-3）,IL-6（mIL-6） and SCF（mSCF） for long-term culture.The results showed that transfection of AML1a into BMMNC enhanced colony formation,colony size of the AML1a group was significantly larger than that of the control group,and the colonies were mainly composed of CFU-E and CFU-GEMM.In the long-term culture,AML1a-transfected BMMNC showed differentiation block,while the control cells were in a more mature stage.It is concluded that AML1a may block the normal hematopoiesis at the stage of primitive progenitors.At the same time,AML1a also enhances the proliferation activity of primitive progenitor cells.

关 键 词：AML1a 骨髓单个核细胞 造血细胞增殖 造血细胞分化 

分 类 号：R331.2[医药卫生—人体生理学]