中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (12): 2118-2125.doi: 10.4103/1673-5374.262592

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

生长阻滞特异性转录因子5有利于神经元和胆碱能神经系统的恢复

  

  • 出版日期:2019-12-15 发布日期:2019-12-15
  • 基金资助:

    中国国际自然科学基金项目（81501133），江苏省博士后研究实践创新项目（KYCX17-1931），南通大学本科创新与创业培训项目（2018150），南通大学自然科学基金预研项目（17ZY19），南通大学杏林学院科研基金项目（2018K131），南通市科技项目（JC2018064）

Long non-coding RNA GAS5 promotes PC12 cells differentiation into Tuj1-positive neuron-like cells and induces cell cycle arrest

He-Yan Zhao, Sheng-Tong Zhang, Xiang Cheng, Hao-Ming Li, Lei Zhang, Hui He, Jian-Bing Qin, Wei-Ye Zhang, Yan Sun, Guo-Hua Jin   

  1. Department of Human Anatomy, the Jiangsu Key Laboratory of Neuroregeneration, Medical School, Nantong University, Nantong, Jiangsu Province, China
  • Online:2019-12-15 Published:2019-12-15
  • Contact: Guo-Hua Jin, PhD, jguohua@ntu.edu.cn.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81501133 (to HML); Postgraduate Research & Practice Innovation Program of Jiangsu Province of China, No. KYCX17-1931 (to HYZ); Undergraduate Innovation and Entrepreneurship Training Project of Nantong University of China, No. 2018150 (to STZ); Pre-research Project of Natural Science Foundation of Nantong University of China, No. 17ZY19 (to HH); Scientific Research Fund Project of Nantong University Xinglin College of China, No. 2018K131 (to HYZ); and Nantong Science and Technology Project of China, No. JC2018064 (to HYZ).

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摘要：

生长阻滞特异性转录因子5是一种抗癌基因，其在肿瘤方面的作用已收到广泛研究，但其在神经疾病中的作用尚有待研究。为探索长链非编码RNA生长阻滞特异性转录因子5对PC12细胞分化的影响，实验将目的基因长链非编码RNA生长阻滞特异性转录因子5过表达慢病毒转染到PC12细胞中，以实时定量PCR分析检测生长阻滞特异性转录因子5以及C-myc mRNA的表达，以EdU检测S期细胞的比例，以免疫组化染色检测神经元微管标志物Tuj1，DCX和微管相关蛋白2的免疫阳性反应，以流式细胞术检测细胞的凋亡，以ELISA分析培养基中乙酰胆碱的表达情况，以Western blot检测细胞中乙酰胆碱转移酶的相对表达情况。(3)结果发现，转染生长阻滞特异性转录因子5可促进PC12细胞分化为具有更长突起的Tuj1阳性神经元样细胞，且细胞增殖和细胞周期受到明显抑制，但不影响细胞凋亡，同时胆碱乙酰转移酶表达和乙酰胆碱释放增加。因此作者推测长链非编码RNA生长阻滞特异性转录因子5有利于神经元和胆碱能神经系统的恢复。

orcid: 0000-0001-7853-2574(Guo-Hua Jin)

关键词: GAS5, PC12细胞, 神经元, 细胞增殖, 细胞周期, 乙酰胆碱转移酶, 乙酰胆碱, 阿尔茨海默病, 神经再生

Abstract:

Growth arrest-specific 5 (GAS5) is an anti-oncogene that has been extensively studied in tumors. However, research on GAS5 in the context of nervous system disease is rare at present. This study aimed to investigate the role of the long non-coding RNA GAS5 in rat pheochromocytoma cells (PC12 cells). GAS5-overexpressing lentivirus was transfected into PC12 cells, and expression levels of GAS5 and C-myc were detected by real-time PCR. Ratios of cells in S phase were detected by 5-ethynyl-2′-deoxyuridine. Immunohistochemical staining was used to detect the immunoreactivity of neuron microtubule markers Tuj1, doublecortin, and microtubule-associated protein 2. Apoptosis was detected by flow cytometry, while expression of acetylcholine in cells was detected by western blot assay. We found that GAS5 can promote PC12 cells to differentiate into Tuj1-positive neuron-like cells with longer processes. In addition, cell proliferation and cell cycle were significantly suppressed by GAS5, whereas it had no effect on apoptosis of PC12 cells. Our results indicate that GAS5 could increase the expression of choline acetyltransferase and acetylcholine release. Thus, we speculate that GAS5 is beneficial to the recovery of neurons and the cholinergic nervous system.

Key words: nerve regeneration, growth arrest-specific 5, PC12 cell, neuron, proliferation, cell cycle, choline acetyltransferase, acetylcholine, Alzheimer’s disease, neural regeneration