miR-29a Modulates Neuronal Differentiation through Targeting REST in Mesenchymal Stem Cells

PLoS One

Ying Xing¹  Shiling Sun²  Chuntian Huang³  Bo Li³  Wenhai Yan³  Ping Duan³  Yan Xu³  Xuefei Han³ 

[1] Department of Physiology, Xinxiang Medical University, Xinxiang, Henan, China;Hematology Department in the First Affiliated Hospital of Henan University of Traditional Chinese Medicine, Zhengzhou, Henan, China;Institute of Basic Medicine, Zhengzhou University, Zhengzhou, Henan, China

关键词: Neuronal differentiation;    MicroRNAs;    Mesenchymal stem cells;    Neuronal morphology;    Gene expression;    Neurons;    Luciferase;    Precursor cells;   

DOI  :  10.1371/journal.pone.0097684

学科分类：医学（综合）

来源: Public Library of Science

【 摘 要 】

Objective To investigate the modulation of microRNAs (miRNAs) upon the neuronal differentiation of mesenchymal stem cells (MSCs) through targeting RE-1 Silencing Factor (REST), a mature neuronal gene suppressor in neuronal and un-neuronal cells.Methods Rat bone marrow derived–MSCs were induced into neuron-like cells (MSC-NCs) by DMSO and BHA in vitro. The expression of neuron specific enolase (NSE), microtubule-associated protein tau (Tau), REST and its target genes, including synaptosomal-associated protein 25 (SNAP25) and L1 cell adhesion molecular (L1CAM), were detected in MSCs and MSC-NCs. miRNA array analysis was conducted to screen for the upregulated miRNAs after neuronal differentiation. TargetScan was used to predict the relationship between these miRNAs and REST gene, and dual luciferase reporter assay was applied to validate it. Gain and loss of function experiments were used to study the role of miR-29a upon neuronal differentiation of MSCs. The knockdown of REST was conducted to show that miR-29a affected this process through targeting REST.Results MSCs were induced into neuron-like cells which presented neuronal cell shape and expressed NSE and Tau. The expression of REST declined and the expression of SNAP25 and L1CAM increased upon the neuronal differentiation of MSCs. Among 14 upregulated miRNAs, miR-29a was validated to target REST gene. During the neuronal differentiation of MSCs, miR-29a inhibition blocked the downregulation of REST, as well as the upregulation of SNAP25, L1CAM, NSE and Tau. REST knockdown rescued the effect of miR-29a inhibition on the expression of NSE and Tau. Meanwhile, miR-29a knockin significantly decreased the expression of REST and increased the expression of SNAP25 and L1CMA in MSCs, but did not significantly affect the expression of NSE and Tau.Conclusion miR-29a regulates neurogenic markers through targeting REST in mesenchymal stem cells, which provides advances in neuronal differentiation research and stem cell therapy for neurodegenerative diseases.

【 授权许可】

CC BY