中间神经元前体细胞株GE6在不同分化条件下小RNA的表达

发布时间：2018-03-15 06:27

本文选题：小RNA　切入点：小RNA芯片　出处：《生物医学工程学杂志》2015年06期 　论文类型：期刊论文

【摘要】：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。
[Abstract]:The purpose of this study was to investigate the expression of microRNAs in the differentiation of rat brain interneuron precursor cell line GE6 in vitro. Small RNAs specifically expressed during the differentiation and maturation of intermediate neuron precursor cell lines were screened and their functions were predicted and analyzed. In the experiment, GE6 was cultured under three different conditions, namely, no addition of any other growth factors, GE6Ge 6d after normal differentiation culture; GE6Ge 6bmp2T after adding bone morphogenetic protein (BMP2) and GE6Ge 6bmp2C after differentiation culture; GE6Ge 6hhhhhhhhhhhhhhhhhhhhhhhhhhhhhhhof differentiation culture 4 days after normal differentiation and culture. The negative control group was divided into undifferentiated GE6Ge 6UU cells. The GE6 cells of the four groups were immunized with cell immunity. The cells of the four groups were immunized with GE6Ge 6U cells. Fluorescence staining qRT-PCR and small RNA microarray analysis were used to compare the four groups of cells. Screening and analysis revealed, There were significant changes in the contents of several small RNA during the differentiation of GE6 cells. MiR-710 miR-290-5p and miR-3473 were significantly up-regulated in GE6 cells added with secretory factor BMP2. These small RNA may play an important role in regulating the differentiation of rat intermediate neurons.
【作者单位】：四川大学华西第二医院发育与干细胞研究所;中国科学院神经科学研究所;
【基金】：国家自然科学基金资助项目(30971633,31171045)
【分类号】：R338;Q78

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