锌转运体7基因沉默抑制APP转基因细胞APP表达和Aβ分泌

ZnT7 gene silencing inhibits the expression of APP and the secretion of Aβ in APP transgenic cells

目的锌转运体7(ZnT7)在阿尔茨海默病(Alzheimer'sdisease,AD)患者脑内高表达并在老年斑内形成聚集效应。但其是否参与AD发病尚不清楚,本实验旨在探讨ZnT7基因沉默对APP转基因细胞APP/Aβ表达水平的影响。方法应用RNAi技术对APP转基因细胞进行ZnT7基因沉默,并采用RT-PCR,WesternBlot,免疫荧光,ELISA,MTT方法,分析ZnT7基因沉默对APP转基因细胞对APP蛋白的表达、Aβ分泌和细胞活力的影响。结果免疫荧光双标结果显示,ZnT7RNAi可明显抑制APP转基因细胞的ZnT7和APP表达;WesternBlot结果显示,ZnT7RNAi可明显抑制APP转基因细胞的APP蛋白表达;ELISA结果显示,ZnT7RNAi可明显抑制APP细胞转基因的Aβ分泌。结论 ZnT7可能参与细胞内APP代谢,ZnT7基因可作为AD治疗的一个潜在靶点。

Objective ZnT7 is expressed in the brain of Alzheimer＇s disease（AD） patients with an aggregation effect in the senile plaques, but it＇s unknown whether it is involved in the development of AD, this experiment is designed to investigate the effects of ZnT7 gene silencing on the expression levels of APP/A β in in APP transgenic cells. Methods RNAi technology was used to silence the ZnT7 gene in the APP transgenic cells, and then RT-PCR,Western Blot, Immunofluorescence, ELISA and MTT were adopted to analyze the effects of ZnT7 gene silencing on the expression of APP, the secretion of A β and the activity of cells in APP transgenic cells. Results Double immunofluorescence and fluorescence microscopy showed that ZnT7 RNAi could significantly inhibit the APP expression level in APP transgenic cells. Western Blot results showed that ZnT7 RNAi could significantly inhibit the APP expression in APP transgenic cells. ELISA results showed that ZnT7 RNAi significantly inhibitted A β secretion. Conclusion ZnT7 may play a role in the intracellular metabolism of APP and ZnT7 gene may be a potential therapeutic target for AD.