Comparative, genome-scale transcriptional analysis of CHRF-288-11 and primary human megakaryocytic cell cultures provides novel insights into lineage-specific differentiation
Comparative, genome-scale transcriptional analysis of CHRF-288-11 and primary human megakaryocytic cell cultures provides novel insights into lineage-specific differentiation
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DOI：
10.1016/j.exphem.2006.10.017
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发表时间：
2007-03-01
影响因子：
2.6
通讯作者：
Papoutsakis, Eleftherios T.
中科院分区：
文献类型：
--
作者：
Fuhrken, Peter G.;Chen, Chi;Papoutsakis, Eleftherios T.
目标.关于巨核细胞分化的转录事件知之甚少。我们试图鉴定以前与巨核细胞生成无关的基因和途径，并评估CHRF-288-11（CHRF）巨核细胞系作为研究巨核细胞生成的模型系统。利用DNA微阵列、Q-RT-PCR和蛋白质水平分析，我们比较了佛波酯诱导CHRF细胞分化和苦参碱诱导人动员外周血CD 34（+）细胞Mk分化的动态基因表达模式。两个系统中已知Mk基因的转录模式相似。CHRF细胞组成型表达一些早期Mk基因，包括加塔-1。凋亡相关基因的表达模式表明，增加p53活性参与MK凋亡，这是证实了p53-DNA结合活性数据和流式细胞术分析的p53靶基因BBC 3。某些Rho和G蛋白偶联受体信号通路组分被上调，包括先前与Mk细胞无关的基因。本体论分析揭示了防御反应基因的上调，包括已知的和候选的血小板衍生的炎症贡献者。干扰素应答基因的上调发生在细胞系中，而不是在原代细胞中，可能是由于JAK 2/STAT 5信号通路中的已知基因突变。这种分析megakaropoiesis，它集成了动态基因表达数据与蛋白质丰度和活性测定，已经确定了一些基因和途径，可能有助于管理megakaropoiesis。此外，转录数据支持CHRF细胞类似于早期Mk表型的假设，并且具有一定的局限性，在用佛波醇酯治疗后表现出真正的Mk分化的转录特征。(c)2007年国际实验血液学学会。爱思唯尔公司出版
Objectives. Little is known about the transcriptional events underlying megakaryocytic (Mk) differentiation. We sought to identify genes and pathways previously unassociated with megakaryopoiesis and to evaluate the CHRF-288-11 (CHRF) megakaryoblastic cell line as a model system for investigating megakaryopoiesis.Methods. Using DNA microarrays, Q-RT-PCR, and protein-level assays, we compared the dynamic gene expression pattern of phorbol ester-induced differentiation of CHRF cells to cytokine-induced Mk differentiation of human mobilized peripheral blood CD34(+) cells.Results. Transcriptional patterns of well-known Mk genes were similar between the two systems. CHRF cells constitutively express some early Mk genes including GATA-1. Expression patterns of apoptosis-related genes suggested that increased p53 activity is involved in Mk apoptosis, and this was confirmed by p53-DNA-binding activity data and flow-cytometric analysis of the p53 target gene BBC3. Certain Rho and G-protein-coupled-receptor signaling pathway components were upregulated, including genes not previously associated with Mk cells. Ontological analysis revealed upregulation of defense-response genes, including both known and candidate platelet-derived contributors to inflammation. Upregulation of interferon-responsive genes occurred in the cell line, but not in the primary cells, likely due to a known genetic mutation in the JAK2/STAT5 signaling pathway.Conclusions. This analysis of megakaryopoiesis, which integrates dynamic gene expression data with protein abundance and activity assays, has identified a number of genes and pathways that may help govern megakaryopoiesis. Furthermore, the transcriptional data support the hypothesis that CHRF cells resemble an early Mk phenotype and, with certain limitations, exhibit genuine transcriptional features of Mk differentiation upon treatment with phorbol esters. (c) 2007 International Society for Experimental Hematology. Published by Elsevier Inc.