miR-124-3p inhibits N2a/APPswe cell apoptosis and cellular calcium ion concentration via down-regulating Caveolin-1

Abstract:

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.