Neural Regeneration Research ›› 2019, Vol. 14 ›› Issue (12): 2118-2125.doi: 10.4103/1673-5374.262592

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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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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