microRNA在胚胎干细胞分化的心肌细胞中的表达变化

发布时间：2018-11-05 12:24

【摘要】：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,P0.05),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,P0.05)。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。
[Abstract]:Embryonic stem cell (embryonic stem cells,ESCs) has the characteristics of self-renewal and multi-differentiation. Recent studies have shown that posttranscriptional regulation, including the regulation of micro RNAs (mi RNAs), plays an important role in determining the fate of cardiomyocyte differentiation (ESC-derived cardiomyocytes,ESCM) in ESC. However, the knowledge of mi RNAs, which determines the fate of ESC differentiation, is still very limited. In order to further understand the regulation of mi RNAs on cardiomyocyte differentiation, the classical suspension method was used to induce ESC (m ESC) to differentiate into cardiomyocytes. Agilent 8 脳 15k mouse mi RNAs chip (mi Rbase V16.0 was used to compare the mi RNAs expression profiles between the regions rich in beating cardiomyocytes and those in non-beating areas. There were more than 5 fold changes in the expression of 19 mi RNAs in the beating region (nang3p0.05), 5 of which were up-regulated and 14 of mi RNAs were down-regulated. Quantitative RT-PCR was used to further analyze the difference ratio of mi RNAs, in mi RNAs chip. The results showed that the expression abundance of mi R-196aanmi R-196b and mi R-467e in the beating region of cardiomyocytes was significantly lower than that in the non-beating region (nnc3p0.05). The target genes of mi R-196a and mi R-196b were predicted by using Target Scan database. It was found that the target genes were negatively correlated with the differentiation of cardiomyocytes, suggesting that they may play a role in regulating the fate of ESC differentiation. These results provide a new clue to further clarify the role of mi RNAs in ESC differentiated cardiomyocytes.
【作者单位】：上海交通大学医学院/中国科学院上海生命科学研究院健康科学研究所中国科学院干细胞生物学重点实验室;
【基金】：supported by the National Basic Research Development Program of China(No.2011CB965301,2014CB965100) the Strategic Priority Research Program of Chinese Academy of Sciences,China(No.XDA01020204,GJHZ1225) the National Science and Technology Major Project of China(No.2012ZX09501001)
【分类号】：R329.2

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