Oct-4在胚胎来源神经前体细胞向神经元分化中的作用

Function of OCT- 4 in differentiation of embryonic neural precursor cells to neurons

目的应用RNA干扰方法抑制胚胎来源神经前体细胞Oct-4基因表达，以观察Oct-4基因在胚胎来源神经前体细胞向神经元分化中的作用。方法采用“五步法”诱导小鼠胚胎干细胞向神经元分化，进入扩增期后给予Oct-4干扰RNA，采用Real-timePCR方法检测干扰后细胞Oct-4表达情况，进行分化计数，免疫组化鉴定分化后的细胞，计数分化后的新生神经元比例。结果Oct-4干扰RNA使神经前体细胞Oct-4表达下降约40％，正常未干扰胚胎干细胞分化的神经元为（75．5±5．2）％，GFP组为（73．8±4．7）％，Oct4干扰RNA组为（33．8±2．9）％，与正常组和GFP组比较，差异均有统计学意义（P〈0．01）。结论采用RNA干扰方法可以有效降低神经前体细胞0ct-4的表达，使分化后新生神经元生成减少，证实了Oct-4基因在胚胎来源神经前体细胞向神经元分化中发挥着重要作用。

Objective Using RNA interference methods to inhibit Oct--4 gene expression in em- bryonic neural precursor cells , to observe the function of OCT--4 in differentiation of embryonic neural precursor cells to neurons. Methods ＂Five step＂ methods were used to induce mouse embryonic stem cells differentiation into neurons, OCT--4 siRNA was given in Amplification stage. Using Real--time PCR method to detect Oct--4 expression in interference cells, identification of the differentiated cells , count the new bron neurons. Results Oct-- 4 RNA interference made neural precursor ceil Oct-- 4 ex- pression decreased about 40%, normal undisturbed embryonic stem cell differentiation neuron about （75. 5±5.2） %, GFP group were（73.8±4.7） % ,Oct4 interference of group RNA neurons were generated about （33.8±2.9）% ,which compared with the control group and GFP group were significantly different （P 〈C 0. 01）. Conclusions Using RNA interference method can effectively reduce the expression of Oct --4 neural precursor cells, reduce of newborn neurons after differentiation, confirmed that the Oct--4 gene plays an important role in embryonic neural precursor cells differentiated into neurons.