Notch-1基因与人神经胶质瘤细胞自噬相关性的研究.pdf

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1、分类号：R34学校代码：10392学科专业代码：071010学号：200901457福建医科大学硕士研究生毕业论文Notch-1基因与人神经胶质瘤细胞自噬相关性的研究ThestudyonthecorrelationbetweenNotch-1geneandautophagyinhumangliomacell学位类型：理学硕士所在学院：基础医学院研究生：阳丽君学科、专业：生物化学与分子生物学导师：郑志竑教授研究起止日期：2010年9月至2012年5月答辩日期：2012年6月2日二○一二年六月万方数据万方数据目录中文摘要···················

2、··································1英文摘要·····················································2英文缩略词表······················································4前言·····················································5实验材料·····················································7方法··········

3、···········································12结果·····················································26讨论·····················································36结论·····················································39参考文献·····················································40致谢···

4、··················································42综述·····················································43万方数据福建医科大学硕士研究生毕业论文Notch-1基因与人神经胶质瘤细胞自噬相关性的研究中文摘要目的：观察上调和下调Notch-1基因表达时人神经胶质瘤细胞的自噬现象，探讨Notch-1基因参与人神经胶质瘤细胞自噬的可能机制。方法：利用表达Notch1-shRNA和Notch-1胞内段（Notch-1intracellulardomain

5、,NICD）的慢病毒感染人胶质瘤U251细胞，通过RT-PCR和WesternBlotting法鉴定Notch-1表达下调或NICD表达上调的U251细胞；应用单丹（磺）酰戊二胺（Monodansylcadaverine，MDC）染色法观察细胞自噬现象；通过Western-blotting和细胞免疫染色法检测自噬相关蛋白LC-3（microtubule-associatedprotein1lightchain3，微管相关蛋白1轻链3）和Beclin-1的表达情况。结果：Notch-1RNA干扰慢病毒能有效下调U251细胞Notch-1的表达，而NICD过

6、表达慢病毒能显著上调U251细胞NICD的表达；MDC荧光染色法发现，Notch-1下调的神经胶质瘤U251细胞中可观察到点状自噬囊泡（autophagicvacuoles，AVs），Western-blotting和细胞免疫染色法结果显示，在Notch-1表达下调的神经胶质瘤U251细胞中，自噬相关蛋白LC-3和Beclin-1表达增高。结论：Notch-1表达下调的人神经胶质瘤细胞可观察到自噬现象，Notch-1表达下调可能导致LC-3和Beclin-1表达上调而诱导胶质瘤细胞自噬。关键词：神经胶质瘤，Notch1信号通路，自噬，LC-3，Becli

7、n-11万方数据福建医科大学硕士研究生毕业论文ThestudyonthecorrelationbetweenNotch-1geneandautophagyinhumangliomacellAbstractObjective:ToinvestigateautophagyphenomenoninhumangliomacellwhentheexpressionofNotch1geneweredown-regulatedorup-regulatedandresearchthepossiblemechanismofNotch-1geneinvolvedinauto

8、phagyinhumangliomacell.Methods:Thelentiv