MicroRNA-1 Regulates Smooth Muscle Cell Differentiation by Repressing Kruppel-Like Factor 4
MicroRNA-1 Regulates Smooth Muscle Cell Differentiation by Repressing Kruppel-Like Factor 4
复制标题
MicroRNA-1 通过抑制 Kruppel 样因子 4 调节平滑肌细胞分化
DOI：
10.1089/scd.2010.0283
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发表时间：
2011-02-01
影响因子：
4
通讯作者：
Chen, Y. Eugene
中科院分区：
文献类型：
--
作者：
Xie, Changqing;Huang, Huarong;Chen, Y. Eugene
MicroRNA-1(miR-1)在心肌和骨骼肌分化中的作用已有研究。然而，它在血管平滑肌细胞(SMCs)分化中的作用尚不清楚。本研究的目的是揭示miR-1在体外胚胎干细胞(ESC)分化中的新靶点并阐明miR-1的功能。在小鼠ESC向SMC分化的过程中，MIR-1的表达持续增加。使用miR-1抑制剂的功能丧失方法发现，miR-1是ESC来源的SMC培养中SMC谱系分化所必需的，SMC特异性标志物的下调和来源SMC数量的减少证明了这一点。此外，生物信息学分析揭示了Kruppel样因子4(KLF4)3‘非翻译区(3’UTR)上的miR-1结合位点，该区域在物种间高度保守。一贯地，miR-1模拟降低的KLF4 3‘UTR荧光素酶活性，这可以通过突变报告构建体中KLF4 3’UTR上的miR-1结合位点来挽救。此外，miR-1抑制剂抑制miR-1的表达可以逆转KLF4在ESC-SMC分化过程中的下调，从而抑制SMC的分化。结论：在维甲酸诱导的ESC/SMC分化过程中，miR-1在决定SMC命运中起着关键作用，提示miR-1可能具有促进SMC分化的作用。
The role of microRNA-1 (miR-1) has been studied in cardiac and skeletal muscle differentiation. However, it remains unexplored in vascular smooth muscle cells (SMCs) differentiation. The aim of this study was to uncover novel targets of and shed light on the function of miR-1 in the context of embryonic stem cell (ESC) differentiation of SMCs in vitro. miR-1 expression is steadily increased during differentiation of mouse ESC to SMCs. Loss-of-function approaches using miR-1 inhibitors uncovered that miR-1 is required for SMC lineage differentiation in ESC-derived SMC cultures, as evidenced by downregulation of SMC-specific markers and decrease of derived SMC population. In addition, bioinformatics analysis unveiled a miR-1 binding site on the Kruppel-like factor 4 (KLF4) 3' untranslated region (3'UTR), in a region that is highly conserved across species. Consistently, miR-1 mimic reduced KLF4 3'UTR luciferase activity, which can be rescued by mutating the miR-1 binding site on the KLF4 3'UTR in the reporter construct. Additionally, repression of the miR-1 expression by miR-1 inhibitor can reverse KLF4 downregulation during ESC-SMC differentiation, which subsequently inhibits SMC differentiation. We conclude that miR-1 plays a critical role in the determination of SMC fate during retinoid acid-induced ESC/SMC differentiation, which may indicate that miR-1 has a role to promote SMC differentiation.