Mechanism of SB431542 in inhibiting mouse embryonic stem cell differentiation
Mechanism of SB431542 in inhibiting mouse embryonic stem cell differentiation
复制标题
SB431542抑制小鼠胚胎干细胞分化的机制
DOI：
10.1016/j.cellsig.2014.06.002
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发表时间：
2014-10-01
影响因子：
4.8
通讯作者：
Guo, Zekun
中科院分区：
文献类型：
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作者：
Du, Juan;Wu, Yongyan;Guo, Zekun
SB 431542（SB）是一种已确立的小分子抑制剂，可特异性结合激活素受体样激酶受体ALK 5、ALK 4和ALK 7的ATP结合结构域，从而特异性抑制Smad 2/3活化并阻断TGF-β信号转导。SB维持小鼠胚胎干细胞的未分化状态。然而，SB维持小鼠胚胎干细胞未分化状态的方式仍不清楚。考虑到当白血病抑制因子退出时SB不能维持胚胎干细胞的多能性，我们试图确定SB维持多能性的机制。通过微阵列和深度序列实验检测SB调控的转录物，包括信使RNA和小的非编码RNA。经检测、Western blot分析和实时荧光定量PCR验证，发现SB调控与自我更新和分化相关的转录本表达。SB主要通过抑制分化发挥作用。SB对关键多能因子的表达无明显影响，但可显著下调成纤维细胞生长因子家族成员等内源性分化相关转录因子的表达。SB可部分抑制维甲酸诱导小鼠胚胎干细胞向神经元分化的反应。(C)2014 Elsevier Inc. All rights reserved.
SB431542 (SB) is an established small molecular inhibitor that specifically binds to the ATP binding domains of the activin receptor-like kinase receptors, ALK5,ALK4 and ALK7, and thus specifically inhibits Smad2/3 activation and blocks TGF-beta signal transduction. SB maintains the undifferentiated state of mouse embryonic stem cells. However, the way of SB in maintaining the undifferentiated state of mouse embryonic stem cells remains unclear. Considering that SB could not maintain embryonic stem cells pluripotency when leukemia inhibitory factor was withdrawn, we sought to identify the mechanism of SB on pluripotent maintenance. Transcripts regulated by SB, including message RNAs and small non-coding RNAs were examined through microarray and deep-sequence experiments. After examination, Western blot analysis, and quantitative real-time PCR verification, we found that SB regulated the transcript expressions related to self-renewal and differentiation. SB mainly functioned by inhibiting differentiation. The key pluripotent factors expression were not significantly affected by SB, and intrinsic differentiation-related transcripts including fibroblast growth factor family members, were significantly down-regulated by SB. Moreover, SB could partially inhibit the retinoic acid response to neuronal differentiation of mouse embryonic stem cells. (C) 2014 Elsevier Inc. All rights reserved.