Let-7d microRNA Attenuates 6-OHDA-Induced Injury by Targeting Caspase-3 in MN9D Cells

Journal of Molecular Neuroscience : MN
Li LiShen Sun

Abstract

Parkinson's disease (PD) mainly results from the progressive loss of dopaminergic (DA) neurons in the substantia nigra pars compacta (SNpc), and the exact underlying mechanisms of the loss of DA neurons in PD remains largely unclear. The results of our previous work showed that let-7d was significantly downregulated in a 6-OHDA-induced cellular model of PD. However, the exact effect of let-7d on DA neural cells was unclear. In MN9D dopaminergic neuronal cells, we used a let-7d mimic and inhibitor to upregulate and downregulate the expression of let-7d, respectively, a cell counting kit to assess cell viability, and a TUNEL staining assay and flow cytometry to examine the cell death rate, and we found that let-7d could negatively regulate 6-OHDA-induced cell injury. Then, we verified that caspase-3 was a target gene of let-7d by using a dual-luciferase reporter system. Furthermore, using caspase-3 siRNA and a caspase-3-overexpression vector (without the 3'UTR) to respectively inhibit and increase the expression of caspase-3, we found that caspase-3 siRNA could reverse the cell injury induced by the let-7d inhibitor and that caspase-3 overexpression could reverse the protective effects of the let-7d mimic on 6-OHDA-induced cell i...Continue Reading

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Citations

Sep 10, 2020·International Journal of Molecular Sciences·Shubhra AcharyaYvan Devaux
Nov 14, 2019·International Journal of Molecular Sciences·Suh Yee GohSamuel Sam-Wah Tay
Oct 12, 2021·Experimental Gerontology·Brian EvansAlexandre de Lencastre

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Methods Mentioned

BETA
flow cytometry
Transfection
electrophoresis
PCR
Assay
protein assay
fluorescence microscopy

Software Mentioned

Statistical Package for the Social Sciences ( SPSS )
TargetScan
ImageJ
miRWalk
miRanda

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