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胚胎干细胞(embryonic stem cells,ESCs)具有自我更新和多向分化的特性。近年研究显示转录后调控包括micro RNAs(mi RNAs)的调控在ESC心肌细胞分化(ESC-derived cardiomyocytes,ESCM)命运决定中起着重要作用。然而对决定ESC分化命运的mi RNAs的了解还非常有限。为了进一步认识mi RNAs对心肌细胞分化的调控作用,本研究采用经典的悬滴法诱导小鼠ESC(m ESC)分化成心肌细胞,采用安捷伦8×15k小鼠mi RNAs芯片(mi Rbase V16.0)比较了富含跳动心肌细胞区域与非跳动区域mi RNAs表达谱的异同,发现与非跳动区域相比,跳动区域中有19个mi RNAs发生了5倍以上的表达变化(n=3,P<0.05),其中5个mi RNAs表达上调、14个mi RNAs表达下调。采用定量RT-PCR进一步分析了mi RNAs芯片中差异倍数大于10的mi RNAs,证明mi R-196a,mi R-196b和mi R-467e在心肌细胞跳动区域的表达丰度明显低于非跳动区域(n=3,P<0.05)。用Target Scan数据库对mi R-196a和mi R-196b进行靶基因预测,发现其可能与心肌细胞分化呈负相关,提示其在ESC分化命运决定中可能具有一定的调控作用。这些结果为进一步明确mi RNAs在ESC分化的心肌细胞中的作用提供了新线索。
Embryonic stem cells (ESCs) have the characteristics of self-renewal and multidirectional differentiation. Recent studies show that post-transcriptional regulation, including the regulation of miRNAs, plays an important role in the fate determination of ESC-derived cardiomyocytes (ESCM). However, the understanding of the mi RNAs that determine the fate of ESC differentiation is still very limited. In order to further understand the regulatory effect of mi RNAs on cardiomyocyte differentiation, we used classical hanging drop method to induce the differentiation of mouse ESCs (m ESC) into cardiomyocytes. Using Agilent 8 × 15k mouse mi RNAs chip (mi Rbase V16.0 ) Compared miRNA expression profiles in beating cardiomyocytes with non-beating regions and found that 19 miRNAs in the beating region were more than 5-fold altered (n = 3, P <0.05). Among them, 5 miRNAs were up-regulated and 14 miRNAs were down-regulated. The miRNAs with mi of multiple of more than 10 in miRNAs chip were further analyzed by quantitative RT-PCR. The results showed that the abundance of mi R-196a, mi R-196b and mi R-467e in cardiomyocytes was significantly lower than that in non-beating area (n = 3, P <0.05). Using Target Scan database to predict mi R-196a and mi R-196b target genes, we found that it may be negatively correlated with cardiomyocyte differentiation, suggesting that it may play a regulatory role in the fate decision of ESC differentiation. These results provide new clues to further clarify the role of mi RNAs in ESC-differentiated cardiomyocytes.