转录因子MZF-1对急性单核细胞白血病新抗原基因MLAA-34的转录调控研究  

Effects of Transcription Factor MZF-1 on Transcriptive Regulation of Acute Monocytic Leukemia-related Gene MLAA-34

作　　者：雷博[1] 张王刚[1] 何爱丽[1] 陈银霞[1] 曹星梅[1] 张鹏宇[1] 赵万红[1] 王剑利[1] 刘捷[1] 马肖容[1] 张彦平[1] 张卉[1] LEI Bo;ZHANG Wang-Gang;HE Ai-Li;CHEN Yin-Xia;CAO Xing-Mei;ZHANG Peng-Yu;ZHAO Wan-Hong;WANG Jian-Li;LIU Jie;MA Xiao-Rong;ZHANG Yan-Ping;ZHANG Hui(Department of Hematology,The Second Affiliated Hospital,Medical School of Xi'an Jiaotong University,Xi'an 710004,Shaanxi Province,China)

机构地区：[1]西安交通大学第二附属医院血液科

出　　处：《中国实验血液学杂志》2019年第5期1463-1468,共6页Journal of Experimental Hematology

基　　金：国家自然科学基金资助(8153000580)

摘　　要：目的:探讨转录因子MZF-1对急性单核细胞白血病新抗原基因MLAA-34的转录调控作用。方法:利用双萤光素酶报告基因检测系统及定点突变技术分析MZF-1对MLAA-34基因启动子转录活性的影响。通过凝胶电泳迁移率变动检测(electrophoretic mobility slift assay,EMSA)和染色质免疫沉淀检测(chromatin immunoprecipitation,Ch IP)实验,验证MZF-1是否与MLAA-34启动子核心区直接特异性结合。构建MZF-1真核表达载体和干涉载体,转染U937细胞,应用RT-PCR和Western blot法检测MLAA-34基因的转录和表达变化。结果:转录因子MZF-1对MLAA-34基因表达具有调控作用,MZF-1结合序列点突变后,相对荧光素酶活性降低(P<0.01)。EMSA和Ch IP实验从细胞内、外水平分别证明MZF-1可与MLAA-34启动子直接结合而发挥调控作用。在过表达试验中,MZF-1的增加可上调MLAA-34的表达(P<0.05);在干涉试验中,MZF-1的降低可下调MLAA-34的表达(P<0.05)。结论:转录因子MZF-1可与MLAA-34基因启动子上的转录调控区结合,并促进急性单核细胞白血病细胞中MLAA-34基因的转录。Objective:To investigate the transcriptional regulation of transcription factor MZF-1 on acute monocytic leukemia-related gene MLAA-34.Methods:The effect of MZF-1 on the transcriptional activity of MLAA-34 gene promoter was analyzed by luciferase reporter gene detection system and site-directed mutation technique.The EMSA and ChIP assay were used to verify whether MZF-1 directly and specifically binds to the core region of MLAA-34 promoter.The over-expression vector and interference vector of MZF-1 were constructed to transfect U937 cells,and RT-PCR and Western blot were used to detect the transcription and expression changes of MLAA-34 gene.Results:The transcription factor MZF-1 had a regulatory effect on MLAA-34 gene expression,and the relative luciferase activity was decreased after MZF-1 binding point mutation(P<0.01).EMSA and ChIP experiments demonstrated that MZF-1 could directly bind to MLAA-34 promoter and play a regulatory role.In the over-expression test,the increase of MZF-1 could up-regulate the expression of MLAA-34(P<0.05).In the interference test,the decrease of MZF-1 could down-regulate the expression of MLAA-34(P<0.05).Conclusion:Transcription factor MZF-1 can bind to the transcriptional regulatory region on the promoter of MLAA-34 gene and promote the transcription of MLAA-34 gene in acute monocytic leukemia.

关 键 词：急性单核细胞白血病 MLAA-34 MZF-1 转录调控 

分 类 号：R733.71[医药卫生—肿瘤]