神经再生素对皮层细胞基因表达影响的基因芯片研究

DNA Microarray Analysis of Gene Expression of Mantle Cell Induced by NRF

目的 :采用基因芯片技术 ,观察神经再生素对体外培养的胚鼠大脑皮层细胞基因表达的影响。方法 :取ICR胚鼠 (15d)大脑皮层细胞 ,无血清培养。实验组加入神经再生素 (2 μg/ml) ,对照组加入等量基础培养液。培养 6h后 ,抽提实验组和对照组皮层细胞总RNA ,经反转录分别用Cy5、Cy3荧光标记成cDNA探针。将荧光标记的cDNA与MGEC 40s基因表达谱芯片杂交 ,芯片扫描、数据处理。结果 :实验组和对照组大脑皮层细胞出现 64条差异性表达的基因。呈现上调的基因有 :钙调素结合蛋白、锌指蛋白激酶、核糖体蛋白等基因 ;下调基因有 :抑制细胞分裂因子等基因。结论 :神经再生素可作用于体外培养的胚鼠大脑皮层细胞 。

AIM:To analyze gene expression of mouse mantle cell induced by nerve regeneration factor (NRF) by DNA microarray. METHOD:Brain mantle cells of embryonic ICR mouse(15 d) were cultured in L15 medium contained NRF(2 μg/ml). Control cells were cultured in L15 medium only. Total RNAs of experimental and control cells were extracted after 6 h. mRNAs were purified. The fluorescent cDNA probes were prepared through reverse transcription. The probes from control cells were labeled with Cy3 dUTP, those from experimental cells with Cy5 dUTP. MGEC 40s DNA microarray was hybridized with the fluorescent probes. The chip was scanned and analyzed. RESULT:There were 69 genes differentially expressed in the cells induced by NRF. Up regulated genes were calcyclin binding protein, zipper protein kinase, ribosomal protein and so on. Down regulated genes were division inhibitory factor and so on. CONCLUSION:NRF could promote brain mantle cells of embryonic mouse growth by gene expression.