南方医科大学学报2017，Vol.37Issue(7)：875-882,8.DOI:10.3969/j.issn.1673-4254.2017.07.04
沉默核仁素可诱导大鼠神经干细胞的分化
Effect of nucleolin silencing on differentiation of rat neural stem cells in vitro and the molecular mechanism
摘要
Abstract
Objective To investigate the effect of nucleolin silencing on the differentiation of rat neural stem cells (NSCs) and the role of Wnt signaling pathway in mediating such effect. Methods Adenovirus vectors expressing small interfering RNA (siRNA) against nucleolin were constructed, verified, and packaged in HEK293A cells. The adenovirus was then transfected into NSCs isolated from neonatal SD rats and the differentiation of the NSCs was examined by detecting the expressions of neuron specific encloase (NSE) and glial fibrillary acidic protein (GFAP) using immunocytochemistry. The expressions of nucleolin, nestin, Wnt3, and β-catenin in the cells were determined with Western blotting. Results Restriction endonuclease and sequencing analysis verified successful construction of the adenoviral vector expressing nucleolin siRNA (nucleolin-siRNA2). Infection of rat NSCs with nucleolin-siRNA2 significantly lowered nucleolin protein expression as compared with that in negative and blank control groups (P<0.05). The percentages of NSE-positive cells and GFAP-positive cells were significantly higher in NSCs infected with nucleolin-siRNA (P<0.01);the infection also resulted in obviously lowered expression of nestin protein and increased expressions of Wnt3 protein andβ-catenin nucleoprotein in the cells. Conclusions Nucleolin silencing by adenovirus-mediated RNA interference induces the differentiation of NSCs into neurons and astrocytes, which is related with the activation of Wnt signaling pathway.关键词
核仁素/神经干细胞/分化/Wnt信号转导通路Key words
nucleolin/neural stem cells/differentiation/Wnt signaling pathway引用本文复制引用
谌崇峰,杨怀才,柳国胜..沉默核仁素可诱导大鼠神经干细胞的分化[J].南方医科大学学报,2017,37(7):875-882,8.基金项目
广东省自然科学基金博士启动项目(S2013040012302) （S2013040012302）
