过表达Sox2基因可增强人骨髓干细胞向神经元分化（英文）

Overexpression of Sox2 gene enhances neuronal differentiation of adult human bone marrow stem cells

目的:研究Sox2对于人骨髓间充质干细胞(hBMSCs)向神经元跨分化能力的影响。方法:利用Western Blot检测hBMSCs中Sox2、Oct4和c-Myc的表达,将携带Sox2和rhGFP编码序列的Tet-On慢病毒感染hBMSCs,通过多西环素(Dox)处理10 d诱导Sox2和rhGFP表达;利用神经诱导培养基诱导感染慢病毒的hBMSCs分化,通过免疫荧光染色技术检测神经元标记物Tuj1的表达;利用Western Blot技术检测Smad 2/3信号通路蛋白在hBMSCs向神经元分化过程中的表达水平。结果:Western Blot结果显示hBMSCs不仅表达Sox2和Oct4,同时还表达c-Myc;光镜下发现Sox2的过表达促进hBMSCs的形态向神经元样细胞转化,同时通过免疫荧光染色发现Tuj1的表达水平增加; Western Blot证实Sox2基因过表达引起hBMSCs中磷酸化Smad 2/3的表达水平降低。结论:Sox2可通过Smad 2/3信号通路增强人骨髓间充质干细胞向神经元分化。

Objective:To study the effects of sex determining region Y box containing gene 2(Sox2)on human bone marrow mesenchymal stem cells(hBMSCs)trans-differentiation into neurons.Methods:The expression of Sox2,Oct,and c-Myc was tested using Western Blot.Tet-On lentivirus carrying Sox2 and rhGFP were infected into hBMSCs and its expression level was regulated by doxycycline(Dox).Then hBMSCs were cultured in the neuronal induction medium for 10 days.The expression of neuronal marker Tuj1 was detected by immunofluorescence staining.The expression level of p Smad 2/3 protein of hBMSCs was detected by Western Blot.Results:Western Blot results confirmed the expression of Sox2,Oct4,and c-Myc in hBMSCs.Over-expression of Sox2 promoted the ability of hBMSCs tans-differentiation into neuron-like cells.Immunofluorescence staining showed that the expression level of Tuj1 increased.Western Blot results also showed that the expression level of p Smad 2/3 decreased in hBMSCs when over-expression of Sox2.Conclusion:Sox2 could enhance neuronal differentiation of hBMSCs through the Smad 2/3 signaling pathway.