贵州医科大学学报

2006, (01) 1-5

[打印本页] [关闭]
本期目录(Current Issue) | 过刊浏览(Past Issue) | 高级检索(Advanced Search)

RNA干扰抑制神经母细胞瘤细胞α7神经型尼古丁受体基因的表达
The Inhibitory Interference of RNA to α7 Neuronal Acetylcholic Receptor Gene Expression in SH-SY5Y Cells

齐晓岚;官志忠;
QI Xiao-lan,GUAN Zhi-zhong(Molecular Biological Key Laboratory of Guiyang Medical College,Guiyang 550004,China)

摘要(Abstract):

目的:利用小分子干扰RNA(RNA i)技术抑制神经母细胞瘤细胞(SH-SY5Y)中α7神经型尼古丁受体(nAChR)基因的表达,并研究α7 nAChR基因表达受抑制后对细胞存活率及脂质过氧化水平的影响。方法:设计并通过分子生物学方法合成α7 nAChR基因特异性小分子干扰RNA(siRNA),转染SH-SY5Y细胞,培养48h后收集细胞,分别应用逆转录聚合酶链反应(RT-PCR)和西印迹方法(W estern-b lotting)检测转染细胞中α7nAChR mRNA及蛋白表达水平的变化,并用1μmol/Lβ淀粉样肽25-35(Aβ25-35)处理siRNA转染细胞,比色法测定脂质过氧化产物含量。四氮基偶氮唑蓝(MTT)方法测定细胞活力。结果:转染siRNA后,与对照组相比,实验组α7 nAChR mRNA及蛋白表达量降低,抑制率分别为52%和36%,脂质过氧化产物丙二醛含量明显增加,MTT法表明实验组细胞活力明显低于对照组,且能增强Aβ的细胞毒性作用。结论:特异性siRNA能有效抑制α7 nAChR基因的表达,α7 nAChR基因的表达的抑制能增加Aβ的神经毒性作用,间接说明α7 nAChR有一定的神经保护作用,其表达减少与阿尔茨海默病的发生有一定关系。
Objective: To inhibit α7 neuronal acetylcholic receptor(α7 nAChR) gene expression in human neuroblastoma cells and study the effects of inhibited expression of α7 nAChR on cell MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) reduction and the level of lipid peroxidation.Methods: The α7 nAChR gene specific siRNAs were designed and synthesized by molecular biological method,and transfected into neuroblastoma cells.The cells were incubated for 48 hours before harvesting.The α7 nAChR mRNA and protein levels inside were monitored with RT-PCR and western blotting.For the measuring of lipid peroxidation and MTT reduction,the transfected neuroblastoma cells were treated with 1μmol/L Aβ_(25-35),and then,spectrophotometry and MTT method were used respectively.Results: Comparing with control group,in experimental group,α7 nAChR mRNA and protein levels decreased with the inhibitory rates of 52% and 36% respectively,and the product of lipid peroxidation increased,and MTT decreased.The α7 nAChR siRNA also increased the toxicity of Aβ_(25-35) on the cultured cells.Conclusions: The α7 nAChR siRNA can specifically inhibit the expression of α7 nAChR gene in neuroblastoma cells.Inhibition of α7 nAChR gene expression can enhance the toxicity of Aβ.α7 nAChR plays an important role in the neuroprotective function and its expression inhibiton may be connected to the pathogenesis of Alzheimer disease.

关键词(KeyWords): 阿尔茨海默病;转染;神经母细胞瘤;逆转录聚合酶链反应;RNA干扰;α7神经型尼古丁受体
Alzheimer disease;neuroblastoma;transfection;reverse transcriptase polymerase chain reaction;RNA interference;α7 neuronal acetylcholic receptor

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金资助项目(30460045)

作者(Author): 齐晓岚;官志忠;
QI Xiao-lan,GUAN Zhi-zhong(Molecular Biological Key Laboratory of Guiyang Medical College,Guiyang 550004,China)

Email:

DOI: 10.19367/j.cnki.1000-2707.2006.01.001

参考文献(References):

文章评论(Comment):

序号(No.) 时间(Time) 反馈人(User) 邮箱(Email) 标题(Title) 内容(Content)
反馈人(User) 邮箱地址(Email)
反馈标题(Title)
反馈内容(Content)
扩展功能
本文信息
服务与反馈
本文关键词相关文章
本文作者相关文章
中国知网
分享