医学毕业nogo.docxVIP

XX大学 毕业论文 Nogo 姓 名： 2014年6月25日 Nogo 作者：武肖娜，李容，李静雯，鞠躬 【关键词】细胞分化 Effects of NogoC on differentiation and ncuritc extension of PC12 cells 【Abstract] AIM: To study the effects of NogoC on the differentiation and neurite extension of PC12 cells. METHODS: The expression of NogoC in NGFtreated PC 12 cells was detected using RTPCR and immunofluorescence. And pEGFPN 1 NogoC, the GFPtaggcd NogoC vector was transfected into PC 12 cells. The effects of NogoC on the differentiation and neurite extension of PC12 cells were further studied by cell counting. RESULTS: A small amount of NogoC mRNA could be detected in NGFtreated PC12 cells; NogoC was found to be expressed in NGF treated PC 12 cells by immunofluorescence; the neurite extension of NGFtreated PC 12 cells transfected with NogoC was significantly increased according to the results of cell counting. CONCLUSION: NogoC was expressed in NGFtreated PC 12 cells, and overexpressed NogoC would facilitate the differentiation and influence the neurite extension of NGF treated PC12 cells. [Keywords] overexpression; NogoC; PC 12 cells; cell differentiation 【摘耍】Fl的：利用PC12细胞研究NogoC和神经元细胞分化及突起生 长的关系.方法：NGF诱导PC12细胞，利用RTPCR及免疫荧光技术检测细胞 内NogoC的表达，并在PC12细胞屮过表达NogoC,通过细胞计数检测NogoC 对突起生氏的影响.结果：①RTPCR检测到NGF诱导的PC12细胞中有少量 NogoC的mRNA分子表达；② 免疫荧光染色观察到NGF诱导的PC12细胞屮 有少量NogoC的表达；③细胞计数观察到转染NogoC后的PC12细胞在NGF 诱导后突起增长百分数相对丁 EGFP组明显增高.结论：NGF诱导的PC12细 胞中有NogoC的表达，同时过表达NogoC可促进NGF诱导下的PC12细胞的分 化以及影响细胞的突起生长. 【关键词】过表达；NogoC； PC12细胞；细胞分化 0引言 Nogo分子是影响成年哺乳动物中枢神经再生最重要的抑制因子之一 [1-2],已证明作为Nogo分子屮最小的选择性剪接体NogoC在屮枢神经系统 (CNS)神经元中表达[3-4].利用RTPCR, Northern Blot等手段证明了 PC12 细胞中表达NogoA, NogoB的mRNA,但RTPCR并没有检测到PC12细胞中冇 NogoC mRNA的表达.用Northern Blot可以检测到NGF诱导后的PC12细胞屮 有少量NogoC mRNA表达[5].这些研究均提示NogoC与神经元的分化相关.我 们以PC12细胞为神经元模型，研究NogoC在神经元细胞的分化及突起生长屮 的作用. 1材料和方法 1」材料 大鼠嗜珞细胞瘤PC12细胞系购自屮国科学院上海细胞生物研究 所.绿色荧光蛋白(GFP)表达载体pEGFPNl购口 Clontech公司；编码人NogoC 的 cDNA 克隆进 pEGFPNl 构建成 pEGFPNlNogoC [4] . DulbeccoT s modified Eagle s medium (DMEM)、胎牛血清、马血清、OPTIMEM、脂质体转染试剂 LipofectamineTM 2000 TRIZAL 等购自