遗传 ›› 2006, Vol. 28 ›› Issue (7): 778-782.

• 研究报告 • 上一篇    下一篇

Sim2基因对PC12细胞分化的影响

孟宪芳, 郑 瑶, 许 强, 沈 洁, 施 静, 彭 彬   

  1. 华中科技大学同济医学院神经生物学系,湖北省神经系统重大疾病重点实验室,武汉430030

  • 收稿日期:2005-06-29 修回日期:2005-10-04 出版日期:2006-07-10 发布日期:2006-07-10
  • 通讯作者: 孟宪芳

Effect of Sim2 gene on the differrentiation of PC12 cells

MENG Xian-Fang, ZHENG Yao, XU Qiang, SHEN Jie, SHI Jing, PENG Bi

  

  1. Department of Neurobiology, Tongji Medical College, Huazhong University of Science and Technology, The Key Laboratory of Nervous System Disease of Hubei Province,Wuhan 430030, China

  • Received:2005-06-29 Revised:2005-10-04 Online:2006-07-10 Published:2006-07-10

摘要: [摘要] 目的 探讨位于Down综合征关键位点的Sim2基因对PC12细胞分化的影响及其机制。 方法 以pcDNA3-mSim2真核表达载体稳定转染PC12细胞,以倒置相差显微镜镜观察PC12细胞神经突起的变化;以RT-PCR方法检测神经元分化相关基因GAP43和Synapsin I mRNA表达水平的变化;流式细胞仪检测GAP43蛋白的表达。 结果 RT-PCR结果显示, pcDNA3-mSim2转染后,mSim2 mRNA表达明显上调;与对照组相比,转染mSim2的PC12细胞突起数量显著减少,长度明显变短;GAP43和Synapsin I mRNA表达水平明显降低(P<0.05);流式细胞仪检测发现,转染mSim2的PC12细胞GAP43蛋白表达水平显著降低(P<0.05)。 结论 Sim2基因可通过影响神经元的分化参与Down综合征的发生。

关键词: :Sim2, PC12细胞, Synapsin I, GAP43

Abstract: [Abstract] Objective To observe the effect of Sim2 gene transfection on the differentiation of PC12 cells, and to explore the role of Sim2 in the pathogenesis of Down syndrome. Methods PC12 cells were transfected with the pcDNA3-mSim2 plasmid by LipofectamineTM reagent and selected with G418. The morphology of PC12 cells was observed by phase-contrast microscope. The mRNA expression of mSim2, GAP43 and Synapsin I was determined by reverse transcription-polymerase chain reaction (RT-PCR). GAP43 protein expression was detected by flow cytometry. Results Abundant mSim2 expression in the pcDNA3-mSim2 transfected PC12 cells was confirmed by RT-PCR, which suggested a successful transfection. In the cells transfected with pcDNA3-mSim2, there were less neuritis, and the length of neurites was shorter. The mRNA expression of GAP43 and Synapsin I was significantly decreased. Upon flow cytometry, the GAP43 protein expression was significantly suppressed by mSim2 transfecton. Conclusion Sim2 may play an important role in the pathogenesis of Down syndrome by influencing the neurite growth and inhibiting the differentiation of neurons.

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