Maintenance of Self-Renewal and Pluripotency in J1 Mouse Embryonic Stem Cells through Regulating Transcription Factor and MicroRNA Expression Induced by PD0325901.
Maintenance of Self-Renewal and Pluripotency in J1 Mouse Embryonic Stem Cells through Regulating Transcription Factor and MicroRNA Expression Induced by PD0325901.
复制标题
通过调节 PD0325901 诱导的转录因子和 MicroRNA 表达维持 J1 小鼠胚胎干细胞的自我更新和多能性
DOI：
10.1155/2016/1792573
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发表时间：
2016
影响因子：
4.3
通讯作者：
Guo Z
中科院分区：
文献类型：
--
作者：
Ai Z;Shao J;Shi X;Yu M;Wu Y;Du J;Zhang Y;Guo Z
胚胎干细胞（ESC）具有无限生长的能力，并在培养中保持其多能性，这种自我更新能力由特定调控基因和表观遗传修饰控制的几个关键分子途径控制。有研究表明，多种表观遗传调控因子，如miRNA和多能性因子，可以紧密整合到分子通路中，并协同维持ESCs的自我更新。然而，小鼠胚胎干细胞在含血清培养基中似乎是异质性的，由于MEK/ERK的自我激活分化信号。因此，为了寻找在MEK/ERK信号通路中建立ESC特性的关键miRNA和关键调控基因，我们对用或不用PD 0325901（PD）（一种众所周知的MEK/ERK信号通路抑制剂）处理的J1 mESC进行微阵列分析和小RNA深度测序，随后进行western blot分析和定量实时PCR验证;我们发现，PD调节与自我更新和分化相关的转录表达，并拮抗视黄酸（RA-）诱导的分化作用。此外，PD可以显着调节在ESC发育中具有关键功能的多种miRNA的表达。总之，我们的研究结果表明，PD可以通过关键调控基因和ES细胞特异性miRNA增强ESC的自我更新能力，从而影响ESC的自我更新和细胞分化。
Embryonic stem cells (ESCs) have the ability to grow indefinitely and retain their pluripotency in culture, and this self-renewal capacity is governed by several crucial molecular pathways controlled by specific regulatory genes and epigenetic modifications. It is reported that multiple epigenetic regulators, such as miRNA and pluripotency factors, can be tightly integrated into molecular pathways and cooperate to maintain self-renewal of ESCs. However, mouse ESCs in serum-containing medium seem to be heterogeneous due to the self-activating differentiation signal of MEK/ERK. Thus, to seek for the crucial miRNA and key regulatory genes that establish ESC properties in MEK/ERK pathway, we performed microarray analysis and small RNA deep-sequencing of J1 mESCs treated with or without PD0325901 (PD), a well-known inhibitor of MEK/ERK signal pathway, followed by verification of western blot analysis and quantitative real-time PCR verification; we found that PD regulated the transcript expressions related to self-renewal and differentiation and antagonized the action of retinoic acid- (RA-) induced differentiation. Moreover, PD can significantly modulate the expressions of multiple miRNAs that have crucial functions in ESC development. Overall, our results demonstrate that PD could enhance ESC self-renewal capacity both by key regulatory genes and ES cell-specific miRNA, which in turn influences ESC self-renewal and cellular differentiation.