ETS-related Transcription Factors ETV4 and ETV5 Are Involved in Proliferation and Induction of Differentiation-associated Genes in Embryonic Stem (ES) Cells
ETS-related Transcription Factors ETV4 and ETV5 Are Involved in Proliferation and Induction of Differentiation-associated Genes in Embryonic Stem (ES) Cells
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DOI：
10.1074/jbc.m115.675595
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发表时间：
2015-09-11
影响因子：
4.8
通讯作者：
Yokota, Takashi
中科院分区：
文献类型：
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作者：
Akagi, Tadayuki;Kuure, Satu;Yokota, Takashi
胚胎干细胞的多能性和自我更新能力受多种转录因子的调控。本研究发现，ets相关转录因子Etv4和Etv5 （Etv4/5）在未分化的ES细胞中特异性表达，抑制Oct3/4可导致Etv4/5下调。在ES细胞中同时缺失Etv4和Etv5（Etv4/5双敲除（dKO））导致扁平的上皮细胞样外观，而在2i培养基（含有两种GSK3和MEK/ERK抑制剂）中形态变为紧凑的菌落。自我更新标记基因（包括Oct3/4和Nanog）的表达水平在野生型和dKO ES细胞之间相似，而细胞周期蛋白依赖性激酶抑制剂（p16/p19、p15和p57）的过度表达会降低Etv4/5 dKO ES细胞的增殖。分化实验显示，Etv4/5 dKO ES细胞衍生的胚状体比对照小，且dKO衍生的胚状体未诱导Fgf5、Sox1、Pax3等外胚层标记基因的表达。微阵列分析显示，Tcf15、Gbx2、Lrh1、Zic3、Baf60c等干细胞相关基因在Etv4/5 dKO ES细胞中被显著抑制。在Etv4/5 dKO ES细胞中人工表达Etv4和/或Etv5可诱导Tcf15和Gbx2的再表达。这些结果表明，Etv4和Etv5可能通过调控Gbx2和Tcf15参与胚胎干细胞的增殖和诱导分化相关基因。
The pluripotency and self-renewal capacity of embryonic stem (ES) cells is regulated by several transcription factors. Here, we show that the ETS-related transcription factors Etv4 and Etv5 (Etv4/5) are specifically expressed in undifferentiated ES cells, and suppression of Oct3/4 results in down-regulation of Etv4/5. Simultaneous deletion of Etv4 and Etv5( Etv4/5doubleknock-out(dKO)) in ES cells resulted in a flat, epithelial cell-like appearance, whereas the morphology changed into compact colonies in a 2i medium (containing two inhibitors for GSK3 and MEK/ERK). Expression levels of self-renewal marker genes, including Oct3/4 and Nanog, were similar between wild-type and dKO ES cells, whereas proliferation of Etv4/5 dKO ES cells was decreased with overexpression of cyclin-dependent kinase inhibitors (p16/p19, p15, and p57). A differentiation assay revealed that the embryoid bodies derived from Etv4/5 dKO ES cells were smaller than the control, and expression of ectoderm marker genes, including Fgf5, Sox1, and Pax3, was not induced in dKO-derived embryoid bodies. Microarray analysis demonstrated that stem cell-related genes, including Tcf15, Gbx2, Lrh1, Zic3, and Baf60c, were significantly repressed in Etv4/5 dKO ES cells. The artificial expression of Etv4 and/or Etv5 in Etv4/5 dKO ES cells induced re-expression of Tcf15 and Gbx2. These results indicate that Etv4 and Etv5, potentially through regulation of Gbx2 and Tcf15, are involved in the ES cell proliferation and induction of differentiation-associated genes in ES cells.