miR-124 promotes proliferation and neural differentiation of neural stem cells through targeting DACT1 and activating Wnt/β-catenin pathways
miR-124 promotes proliferation and neural differentiation of neural stem cells through targeting DACT1 and activating Wnt/β-catenin pathways
复制标题
miR-124 通过靶向 DACT1 和激活 Wnt_β-catenin 通路促进神经干细胞的增殖和神经分化
DOI：
10.1007/s11010-018-3367-z
复制
发表时间：
2018-12-01
影响因子：
4.3
通讯作者：
Teng，Junfang
中科院分区：
文献类型：
--
作者：
Jiao，Shujie;Liu，Yaling;Teng，Junfang
神经干细胞（Neural stem cells，NSCs）是一种具有向神经元和胶质细胞分化潜能的多能性未分化细胞。神经干细胞具有生成和再生大脑的能力，这表明了基于细胞的神经系统疾病治疗的可能性。miR-124已被证明是NSC存活、扩增和分化的调节剂。然而，miR-124在NSC发育中的潜在分子机制仍远未被理解。采用qRT-PCR或western blot检测miR-124、dishevelled binding antagonist of beta-catenin 1（DACT 1）、ki-67、Nestin、β-tubulin III、胶质细胞酸性蛋白（GFAP）、β-catenin、cyclinD 1和糖原合成酶激酶-3 β（GSK-3β）的表达。利用生物信息学和双荧光素酶报告基因分析技术研究miR-124与DACT 1的相互作用。进行MTS分析以测量NSC的活力。在14天的NSC分化期间观察到miR-124的表达增强和DACT 1的表达降低。DACT 1被证实是miR-124的直接靶点。此外，miR-124过表达促进NSC增殖并诱导神经元特异性分化，表现为细胞活力增加，神经球数量增加，ki-67、Nestin、β-tubulin III表达升高，GFAP表达降低。类似地，DACT 1下调促进NSC的增殖和神经元分化。此外，DACT 1过表达损害了miR-124诱导的NSC增殖和分化。此外，miR-124通过抑制DACT 1表达刺激Wnt/β-catenin信号传导。miR-124通过靶向DACT 1激活Wnt/β-catenin通路，促进NSC增殖并诱导其分化为神经元，为神经系统疾病的细胞治疗提供了潜在的靶点。
Neural stem cells (NSCs) are multipotent and undifferentiated cells with the potential to differentiate into neuronal lineages and gliocytes. NSCs have the ability to generate and regenerate the brain, indicating the possibility of cell-based therapies for neurological disorders. miR-124 has been demonstrated as a modulator in the survival, expansion, and differentiation of NSCs. However, the underlying molecular mechanisms of miR-124 in NSC development are still far from being understood. The expressions of miR-124, dishevelled binding antagonist of beta-catenin 1 (DACT1), ki-67, Nestin, β-tubulin III, glial fibrillary acidic protein (GFAP), β-catenin, cyclinD1, and glycogen synthase kinase-3β (GSK-3β) were examined by qRT-PCR or western blot. Bioinformatics and Dual-Luciferase reporter assay were used to identify the interaction between miR-124 and DACT1. MTS analysis was performed to measure the viability of NSCs. Enhanced expression of miR-124 and lowered expression of DACT1 were observed during a 14-day NSC differentiation period. DACT1 was verified as a direct target of miR-124. Moreover, overexpression of miR-124 promoted NSC proliferation and induced neuron-specific differentiation, presented as increased cell viability, higher neurosphere number, elevated ki-67, Nestin, β-tubulin III expressions, and decreased GFAP expression. Similarly, DACT1 downregulation facilitated proliferation and neuronal differentiation of NSCs. Furthermore, DACT1 overexpression impaired miR-124-induced proliferation and differentiation of NSCs. Additionally, miR-124 stimulated Wnt/β-catenin signaling via suppressing DACT1 expression. miR-124 promoted proliferation and induced NSC differentiation to neurons by activation of Wnt/β-catenin pathway via targeting DACT1, providing a potential target and aiding the development of cell-based therapies for neurological disorders.