内科理论与实践2013，Vol.8Issue(3)：192-196,5.DOI:10.3969/j.issn.1673-6087.2013.03.009
AML1-EVI1转基因斑马鱼模型的建立及对造血转录因子的调控
The establishment of AML1-EVI1 transgenic zebrafish model and regulation of hematopoietic transcription factors
摘要
Abstract
Objective To study the influence of acute myeloid leukemia (AML)1-ecotropic virus integration site (EVI)1 fusion gene on the transcriptional regulation of hematopoiesis by establishment of AML1-EVI1 transgenic zebrafish model.Methods AML1-EVI1 plasmid with reporter gene carrying enhanced green fluorescent protein (EGFP) was constructed.Then the vector was microinjected into single-cell zebrafish embryos to establish the model of stable germline transgenic zebrafish line,which was bred to F2 generation.AML1-EVI1 fusion gene in transgenic zebrafish was detected by RT-PCR.Blood cell morphology was detected by peripheral blood smear and kidney cell smear,and the influence on hematopoietic regulatory factors [globin transcription factor 1 (GATA1),myeloperoxidase (MPO)] was assessed by wholemount in situ hybridization and real time quantitative PCR (RFQ-PCR).Results Three of 146(2.1％) mosaic F0 zebrafish embryos were identified as the positive germline transgenic zebrafish,including 2 males and 1 female.With F2 generation of transgenic zebrafish as experimental object,the morphology of peripheral blood smear and kidney cell smear showed the presence of blast cells and disturbance in differentiation.Whole-mount in situ hybridization and RFQ-PCR displayed that the expression level of MPO gene in the transgenic group was markedly decreased,and the expression level of GATA1 in the transgenic group was increased in the early stage then decreased in the late stage of hematopoiesis in transgenic zebrafish.Conclusions AML1-EVI1 fusion gene could significantly inhibit the erythropoiesis,suppress the myelocyte differentiation and promote primitive cell proliferation by regulation of transcription factors.关键词
AML1-EVI1/斑马鱼模型/造血转录因子Key words
AML1-EVI1/Zebrafish model/Transcription factors分类
医药卫生引用本文复制引用
林文洁,沈莉菁,陈芳源,钟敏,孙瑞林,钟济华..AML1-EVI1转基因斑马鱼模型的建立及对造血转录因子的调控[J].内科理论与实践,2013,8(3):192-196,5.基金项目
国家自然科学基金(项目编号:30900636) （项目编号:30900636）
上海市科委基础研究重点项目(项目编号:08JC1414900) （项目编号:08JC1414900）
上海市卫生局重大课题(中医药)(项目编号:ZYSNXD-CC-ZDYJ001) （中医药）
