中国神经再生研究(英文版) ›› 2020, Vol. 15 ›› Issue (6): 1133-1139.doi: 10.4103/1673-5374.270417

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

miR-124在调节视黄酸诱导N2a细胞分化中的作用

  

  • 出版日期:2020-06-15 发布日期:2020-07-05
  • 基金资助:
    上海市自然科学基金(16ZR1410500)

Role of miR-124 in the regulation of retinoic acid-induced Neuro-2A cell diferentiation

Qun You1 , Qiang Gong 1 , Yu-Qiao Han1 , Rou Pi 1 , Yi-Jie Du2, 3 , Su-Zhen Dong 1   

  1. 1 Shanghai Engineering Research Center of Molecular Terapeutics and New Drug Development, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai, China
    2 Department of Integrative Medicine, Huashan Hospital, Fudan University, Shanghai, China
    3 Institutes of Integrative Medicine, Fudan University, Shanghai, China
  • Online:2020-06-15 Published:2020-07-05
  • Contact: Yi-Jie Du, PhD,duyijie@huashan.org.cn; Su-Zhen Dong, PhD,szdong@brain.ecnu.edu.cn.
  • Supported by:
    This work was supported by the Natural Science Foundation of Shanghai of China, No. 16ZR1410500 (to SZD).

摘要:

视黄酸能诱导包括小鼠成神经瘤细胞N2a在内的许多类型的细胞分化成神经元,目前仍不清楚miRNA在视黄酸诱导N2a细胞分化过程中的作用。为此,实验利用定量PCR检测视黄酸诱导N2a细胞1-5 d后一些分化相关miRNA的表达变化,(1)发现在视黄酸诱导分化的N2a细胞中,miR-124和miR-9表达上调,miR-125b表达下调;(2)为了进一步确定N2a诱导分化过程中起关键作用的miRNA,在N2a细胞中分别转染miR-124的类似物或抑制剂调控其表达,发现抑制miR-124表达能减少视黄酸诱导的神经突生长,单独miR-124过表达即诱导N2a细胞分化成神经元,同时转染mi-124抑制剂能阻断该效果;(3)上述结果表明,miR-124在视黄酸诱导N2a细胞分化过程中起重要作用。

orcid: 0000-0003-0619-4371 (Yi-Jie Du)
         0000-0002-6716-6115 (Su-Zhen Dong)

关键词: 视黄酸, 小鼠成神经瘤细胞N2a, 神经元分化, miR-124, 神经突生长, 微管相关蛋白-2, 小RNA, 实时定量PCR, 免疫荧光, 过表达

Abstract: Retinoic acid can cause many types of cells, including mouse neuroblastoma Neuro-2A cells, to diferentiate into neurons. However, it is still unknown whether microRNAs (miRNAs) play a role in this neuronal diferentiation. To address this issue, real-time polymerase chain reaction assays were used to detect the expression of several diferentiation-related miRNAs during the diferentiation of retinoic ac- id-treated Neuro-2A cells. Te results revealed that miR-124 and miR-9 were upregulated, while miR-125b was downregulated in retinoic acid-treated Neuro-2A cells. To identify the miRNA that may play a key role, miR-124 expression was regulated by transfection of miRNA mimics or inhibitors. Morphological analysis results showed that inhibition of miR-124 expression reversed the efects of retinoic acid on neurite outgrowth. Moreover, miR-124 overexpression alone caused Neuro-2A cells to diferentiate into neurons, and its inhibitor could block this efect. Tese results suggest that miR-124 plays an important role in retinoic acid-induced diferentiation of Neuro-2A cells.

Key words: immunofuorescence, MAP2, microRNA, miR-124, Neuro-2A cells, neurite outgrowth, neuronal diferentiation, overexpression, real-time PCR, retinoic acid