MicroRNA-1 and MicroRNA-133 in Spontaneous Myocardial Differentiation of Mouse Embryonic Stem Cells
MicroRNA-1 and MicroRNA-133 in Spontaneous Myocardial Differentiation of Mouse Embryonic Stem Cells
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DOI：
10.1253/circj.cj-08-1032
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发表时间：
2009-08-01
影响因子：
3.3
通讯作者：
Hasegawa, Koji
中科院分区：
文献类型：
--
作者：
Takaya, Tomohide;Ono, Koh;Hasegawa, Koji
背景：MicroRNA (miRNA) 通过抑制 RNA 转录本的翻译来调节各种生物过程。尽管 miRNA-1 (miR-1) 和 miRNA-133 (miR-133) 在成体心脏中大量表达并参与心脏肥大，但这些 miRNA 在自发心肌分化中的作用尚不清楚。 方法和结果：小鼠胚胎干 (ES) 细胞中 miR-1 和 miR-133 的水平在二维培养的自发分化过程中增加，但在组蛋白脱乙酰酶抑制剂曲古抑菌素 A 强制心肌分化过程中降低。在 ES 细胞分化过程中，慢病毒感染引起的 miR-1 或 miR-133 过度表达降低了心脏特异性基因 Nkx2.5 的表达。此外，miR-1还抑制α-肌球蛋白重链表达。荧光素酶检测结果显示，miR-1 识别并靶向 ES 细胞中细胞周期蛋白依赖性激酶 9 (Cdk9) 的 3' 非翻译区。 miR-1的过表达降低了Cdk9的蛋白含量，但不影响mRNA水平，表明miR-1转录后抑制Cdk9翻译。结论：miR-1和miR-133可能在小鼠ES细胞的心肌分化中发挥重要作用，Cdk9可能作为miR-1的靶标参与这一过程。 （Circ J 2009；73：1492-1497）
Background: MicroRNAs (miRNAs) regulate various biological processes through inhibiting the translation of RNA transcripts. Although miRNA-1 (miR-1) and miRNA-133 (miR-133) are abundantly expressed in the adult heart and involved in cardiac hypertrophy, the roles of these miRNAs in spontaneous myocardial differentiation are unknown.Methods and Results: The levels of miR-1 and miR-133 in mouse embryonic stem (ES) cells were increased during spontaneous differentiation by 2-dimensional culture, but reduced during forced myocardial differentiation by a histone deacetylase inhibitor, trichostatin A. The overexpression of miR-1 or miR-133 by lentiviral infection reduced the expression of a cardiac-specific gene, Nkx2.5, during differentiation of ES cells. In addition, miR-1 also inhibited a-myosin heavy chain expression. The results of luciferase assays revealed that miR-1 recognizes and targets the 3' untranslated region of cyclin-dependent kinase-9 (Cdk9) in ES cells. Overexpression of miR-1 decreased the protein amounts of Cdk9 without affecting the mRNA levels, indicating that miR-1 post-transcriptionally inhibits Cdk9 translation.Conclusions: miR-1 and miR-133 may play significant roles in the myocardial differentiation of mouse ES cells, and Cdk9 may be involved in this process as a target of miR-1. (Circ J 2009; 73: 1492-1497)