Gažová Iveta, Lefevre Lucas, Bush Stephen J, Clohisey Sara, Arner Erik, de Hoon Michiel, Severin Jessica, van Duin Lucas, Andersson Robin, Lengeling Andreas, Hume David A, Summers Kim M
The Roslin Institute, The University of Edinburgh, Edinburgh, United Kingdom.
RIKEN Center for Integrative Medical Sciences, Kanagawa, Yokohama, Japan.
Front Cell Dev Biol. 2020 Jul 3;8:498. doi: 10.3389/fcell.2020.00498. eCollection 2020.
The response of the human acute myeloid leukemia cell line THP-1 to phorbol esters has been widely studied to test candidate leukemia therapies and as a model of cell cycle arrest and monocyte-macrophage differentiation. Here we have employed Cap Analysis of Gene Expression (CAGE) to analyze a dense time course of transcriptional regulation in THP-1 cells treated with phorbol myristate acetate (PMA) over 96 h. PMA treatment greatly reduced the numbers of cells entering S phase and also blocked cells exiting G2/M. The PMA-treated cells became adherent and expression of mature macrophage-specific genes increased progressively over the duration of the time course. Within 1-2 h PMA induced known targets of tumor protein p53 (TP53), notably , followed by gradual down-regulation of cell-cycle associated genes. Also within the first 2 h, PMA induced immediate early genes including transcription factor genes encoding proteins implicated in macrophage differentiation () and down-regulated genes for transcription factors involved in immature myeloid cell proliferation (). The dense time course revealed that the response to PMA was not linear and progressive. Rather, network-based clustering of the time course data highlighted a sequential cascade of transient up- and down-regulated expression of genes encoding feedback regulators, as well as transcription factors associated with macrophage differentiation and their inferred target genes. CAGE also identified known and candidate novel enhancers expressed in THP-1 cells and many novel inducible genes that currently lack functional annotation and/or had no previously known function in macrophages. The time course is available on the ZENBU platform allowing comparison to FANTOM4 and FANTOM5 data.
人类急性髓系白血病细胞系THP-1对佛波酯的反应已得到广泛研究，用于测试候选白血病治疗方法，并作为细胞周期停滞和单核细胞-巨噬细胞分化的模型。在此，我们采用基因表达的帽分析（CAGE）来分析用佛波酯肉豆蔻酸酯（PMA）处理96小时的THP-1细胞中转录调控的密集时间进程。PMA处理大大减少了进入S期的细胞数量，也阻止了细胞退出G2/M期。经PMA处理的细胞变得贴壁，成熟巨噬细胞特异性基因的表达在整个时间进程中逐渐增加。在1-2小时内，PMA诱导了肿瘤蛋白p53（TP53）的已知靶标，特别是 ，随后细胞周期相关基因逐渐下调。同样在最初的2小时内，PMA诱导了即时早期基因，包括编码与巨噬细胞分化有关的蛋白质的转录因子基因（ ），并下调了参与未成熟髓样细胞增殖的转录因子基因（ ）。密集的时间进程表明，对PMA的反应不是线性和渐进的。相反，基于网络的时间进程数据聚类突出了编码反馈调节因子以及与巨噬细胞分化相关的转录因子及其推断的靶基因的基因表达的瞬时上调和下调的顺序级联。CAGE还鉴定了在THP-1细胞中表达的已知和候选新型增强子，以及许多目前缺乏功能注释和/或在巨噬细胞中以前没有已知功能的新型诱导基因。该时间进程可在ZENBU平台上获取，以便与FANTOM4和FANTOM5数据进行比较。
