1. [Effects of Transcription Factor MZF-1 on Transcriptive Regulation of Acute Monocytic Leukemia-related Gene MLAA-34]
- Author
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Bo, Lei, Wang-Gang, Zhang, Ai-Li, He, Yin-Xia, Chen, Xing-Meim, Cao, Peng-Yu, Zhang, Wan-Hong, Zhao, Jian-Li, Wang, Jie, Liu, Xiao-Rong, Ma, Yan-Ping, Zhang, and Hui, Zhang
- Subjects
Gene Expression Regulation, Neoplastic ,Transcription, Genetic ,Antigens, Neoplasm ,Genes, Reporter ,Leukemia, Monocytic, Acute ,Kruppel-Like Transcription Factors ,Humans ,Hepatocyte Nuclear Factor 1-alpha ,Apoptosis Regulatory Proteins ,Promoter Regions, Genetic - Abstract
To investigate the transcriptional regulation of transcription factor MZF-1 on acute monocytic leukemia-related gene MLAA-34.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.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 (P0.01). 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 (P0.05). In the interference test, the decrease of MZF-1 could down-regulate the expression of MLAA-34 (P0.05).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.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.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.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,ChIP)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.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(P0.01)。EMSA和ChIP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(P0.05);在干涉试验中,MZF-1的降低可下调MLAA-34的表达(P0.05).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.
- Published
- 2019