C6胶质瘤细胞诱导神经干细胞分化的初步研究

Experimental study on differentiation of neural stem cells induced by C6 glioma cells

为了探讨C6胶质瘤细胞分泌的因子对神经干细胞分化的作用及机制。本研究从Wistar孕鼠(E17～E18)纹状体组织中分离、培养神经干细胞,将其分离纯化后进行nestin免疫荧光染色鉴定后分为两组进行诱导分化实验。对照组:RPMI1640培养液+基础培养基(1∶1配制);处理组:C6胶质瘤细胞培养上清+基础培养基(1∶1配制)。采用免疫印迹法分析胶质瘤细胞条件培养液作用于神经干细胞后的不同时相(1、3、5d)MAP-2蛋白的表达情况;通过RT-PCR检测不同时相(1、3、5d)MAP-2的表达及调控神经干细胞向神经元分化的多种转录因子如:neurogenin3(Ngn3)、neuroD和neuroD2的mRNA的表达情况。结果显示:处理组MAP-2在蛋白和mRNA水平的表达量均明显高于对照组(P<0.01),并且处理组的Ngn3、neuroD和neuroD2的mRNA水平比对照组也有明显升高。上述结果提示,C6胶质瘤细胞条件培养液能够促进神经干细胞向神经元分化,此作用可能与Ngn3、neuroD和neuroD2等转录调控因子表达水平的提高有关。

To explore the effect and mechanism of C6 glioma cells derived factors on the differentiation of neural stem cells. The neural stem cells derived from fetal rat brain were isolated, purified and then were treated with C6 glioma cells conditioned medium in vitro. The level of MAP-2 expression was detected by Western blot analysis and RT-PCR. RT-PCR was also performed to evaluate mRNA levels of multiple transcriptional factors such as neurogenin3 （Ngn3）, neuroD and neuroD2 which are neural stem cells differentiation regulators. The results showed that both protein and mRNA level of MAP-2 in treated group were higher than those of control significantly （P 〈 0. 05 ） , furthermore mRNA level of Ngn3, neuroD and neuroD2 in treated group were also increased compared with control. The above results suggest that C6 glioma cells derived factors can promote neural stem cells differentiate into neurons, which probably due to up-regulation of neural stem cells differentiation transcriptional factors including Ngn3, neuroD and neuroD2.