Xiang Yue, Zhang Xiong, Yang Jun, et al. miR-124-3p inhibits N2a/APPswe cell apoptosis and cellular calcium ion concentration via down-regulating Caveolin-1[J]. Journal of Chongqing Medical University, 2015,40(5):671-676.
Xiang Yue, Zhang Xiong, Yang Jun, et al. miR-124-3p inhibits N2a/APPswe cell apoptosis and cellular calcium ion concentration via down-regulating Caveolin-1[J]. Journal of Chongqing Medical University, 2015,40(5):671-676.DOI:
Objective:To investigate the effect of miR-124-3p on the apoptosis and the concentration of free calcium in cell by regu-lating the expression of Caveolin-1 in Alzheimer’s diseases(AD) cell model of N2a/APPswe cells. Methods:N2a/APPswe and wild type N2a(N2a/WT) cells were cultured in vitro;real-time fluorescence quantitative PCR(real-time PCR) and Western blot(WB) were re-spectively used to detect the expression levels of miR-124-3p and amyloid precursor protein(APP) mRNA and protein. Dual luciferase report experiments were used to detect targeting regulation relationship between miR-124-3p and Caveolin-1. N2a/APPswe cells were transiently transfected by miR-124-3p mimics;real-time PCR and WB were respectively used to detect the expression levels of Caveolin-1 mRNA and protein. N2a/APPswe cells were transiently transfected by miR-124-3p mimics,pcDNA-Caveolin-1,Caveolin-1-siRNA and the rate of cell apoptosis was analyzed by flow cytometry. The intracellular free calcium concentration was measured. Results:Compared with those of WT group,APP mRNA and protein levels were significantly increased(P=0.000,P=0.000) in APP-swe group,miR-124-3p expression was obviously decreased(P=0.000). Dual luciferase report experiment showed that compared with those of co-transfection with the mutant vector(pGL3- Caveolin-1 3’UTR MUT) group,relative luciferase activities in co-transfection with the wild type vector(pGL3-Caveolin-1 3’UTR WT) group were significantly decreased(P=0.004). After the transfection of miR-124-3p mimics,Caveolin-1 mRNA and protein levels were significantly decreased(P=0.000,P=0.000) in transfection group than in control group. After the transfection of miR-124-3p mimics and Caveolin-1-siRNA,the cell apoptosis rate and free calcium concentration were decreased(P=0.000) in transfection group than in control group(P=0.000). After the transfection of pcDNA-Caveolin-1,the opposite results were obtained(P=0.000,P=0.000). Conclusion:miR-124-3p can inhibit the apoptosis of cell and reduce the concentration of free calcium in cell by targeted down-regulation of Caveolin-1 expression,which plays a neuro-protective role in prevention and cure of AD and provides new ideas and targets for AD.
关键词
miR-124-3pCaveolin-1N2a/APPswe细胞凋亡钙离子浓度
Keywords
miR-124-3pCaveolin-1N2a/APPswe cellapoptosiscalcium ion concentration