NGF刺激PC12细胞神经分化过程中转录因子E2F1参与PTPRO表达调控的研究

发布时间：2017-12-28 11:32

本文关键词：NGF刺激PC12细胞神经分化过程中转录因子E2F1参与PTPRO表达调控的研究　出处：《东北师范大学》2009年硕士论文　论文类型：学位论文

更多相关文章： PTPRO E2F1 转录调控 轴突生长

【摘要】： PTPRO是受体型酪氨酸磷酸酯酶家族的一员,其功能主要涉及神经组织中轴突的生长和导向,以及肿瘤抑制。PTPRO的表达具有组织特异性,在哺乳动物中PTPRO主要表达于不同发育时期的肾脏组织和神经组织,而在很多肿瘤细胞中则通过DNA甲基化沉默。有文献报道,PTPRO在机体发育和脑的成熟中发挥重要作用,例如在肾脏中,PTPRO表达于发育早期的肾小球细胞顶部和发育成熟的肾小球足细胞顶部;在脑中,PTPRO基因最高表达出现于妊娠中后期,这一时期也是脑部轴突发生的主要时期。已有研究证明,受体型酪氨酸磷酸酯酶家族(RPTPs)成员在调节果蝇神经系统发育过程中起主要作用。在脊椎动物中,许多受体型酪氨酸磷酸酯酶家族(RPTPs)成员例如:typeⅡa RPTPs,typeⅢRPTP已经在轴突生长过程中发挥重要作用,但是PTPRO参与轴突生长的具体机制以及表达调控机制却报道很少。本论文旨在研究PTPRO基因在NGF诱导PC12轴突分化过程中表达升高的转录调控机制。通过反转录PCR(RT-PCR)、real-time RCR和Western blotinging检测我们发现,NGF诱导PC12轴突分化后,PTPRO基因表达发生了明显的上调。细胞转染和荧光素酶报告基因分析结果显示,外源过表达转录因子E2F1能显著提高PTPRO启动子荧光素酶报告基因的活性。real-time RCR和western bloting结果表明,转录因子E2F1提高了细胞内源PTPTO基因的mRNA和蛋白表达水平。点突变实验确定了PTPRO启动子上E2F1的结合位点。而ChIP实验表明E2F1能够结合在PTPRO的启动子上。通过以上实验,我们初步地研究了PC12细胞在NGF刺激后PTPRO表达上调的调控机制。为下一步更详细的研究PTPRO在神经分化过程中的具体分子机制以及PTPRO基因调控的具体机制定了基础。
[Abstract]:PTPRO is a member of the receptor tyrosine phosphatase family, whose function mainly involves the growth and guidance of axons in the nerve tissue and tumor inhibition. The expression of PTPRO is tissue-specific. In mammals, PTPRO is mainly expressed in different developmental stages of renal tissue and nerve tissue, while in many tumor cells, DNA methylation is silent. There are reports in the literature, PTPRO play an important role in the development of the body and brain maturation, for example in the kidney, the expression of PTPRO in the development of glomerular cells and top early podocyte top mature development; in the brain, the highest expression of PTPRO gene in late pregnancy, the main period of this period is also the occurrence of brain axons. It has been shown that the member of the type tyrosine phosphatase family (RPTPs) plays a major role in regulating the development of the nervous system in Drosophila. In vertebrates, many receptor tyrosine phosphatase family (RPTPs) members, such as type II a RPTPs, type III RPTP, have played an important role in the process of axonal growth. However, there are few reports about the mechanism of PTPRO involved in axonal growth and the regulation mechanism of its expression.
【学位授予单位】：东北师范大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R33;R730.2

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