中间神经元前体细胞株GE6在不同分化条件下小RNA的表达

Expression of MicroRNAs of An Interneuron Precursor Cell Line GE6 in Various Differentiation Conditions

本研究通过分析小RNA(microRNAs)在大鼠大脑中间神经元前体细胞株GE6体外分化中的表达,筛选出在中间神经元前体细胞系分化成熟过程中特异性表达的小RNA,并预测分析其可能在此过程中的功能。实验中,将GE6在三种不同条件下培养,即:不加入任何其他生长因子,正常分化培养4天的GE6(Ge6_4d);加入骨形态发生蛋白-2(BMP2)后分化培养4天的GE6(Ge6_bmp2);以及加入音猥因子(SHH)后分化培养4天的GE6(Ge6_shh)。设阴性对照组为未分化的GE6(Ge6_u)。四组GE6细胞进行细胞免疫荧光染色、qRT-PCR以及小RNA芯片检测分析。通过四组细胞的两两比较、筛选及分析发现,多种小RNA在GE6细胞的分化过程中含量有明显变化。miR-710、miR-290-5p和miR-3473等在加入分泌因子BMP2的GE6细胞中含量显著上调。这些小RNA可能对大鼠中间神经元分化起到重要的调控作用。

The purpose of this study was to identify specific microRNAs （miRNAs） during differentiation and matu- ration of interneurons and to predict their possible functions by analyzing the expression of miRNAs during in vitro differentiation of the rat interneuron precursor cell line GE6. In the experiment, the interneuron precursor cell line GE6 was cultured under three different conditions, i.e. the first was that had not added growth factors and the nor- real differentiation cultured for 4 days （Ge6_4d） ; the second was that cultured with bone morphogenetic protein-2 （BMP2） for 4 days （Ge6_bmp2）; and the third was that cultured with sonic hedgehog （SHH） for 4 days （Ge6_ shh）. In addition, another group of undifferentiated GE6 （Ge6_u） was applied as a control. We found in this study that the expression levels of a large number of miRNAs changed slgn^ficantly during GE6 differentiation. The expres- sion levels of miR-710, miR-290-5p and miR-3473 increased in the GE6 ceils with secreted factor BMP2. These miR- NAs may play important regulatory roles during interneuron differentiation.