Sodium valprovate suppresses autophagy in SH-SY5Y cells activating miR-34c-5p/ATG4B signaling pathway.
10.12122/j.issn.1673-4254.2018.12.03
- Author:
Xufang DAI
1
;
Xiaojing YAN
2
;
Peng XIE
2
;
Jiqin LIAN
2
Author Information
1. Chongqing Key Laboratory of Psychological Diagnosis and Educational Technology for Children with Special Needs.
2. Department ofBiochemistry and Molecular Biology, Army Medical University, Chongqing 400038, China.
- Publication Type:Journal Article
- Keywords:
ATG4B;
SH-SY5Y cells;
autophagy;
miR-34c-5p;
sodium valproate
- MeSH:
Autophagy;
drug effects;
Autophagy-Related Proteins;
genetics;
metabolism;
Cell Line;
Cysteine Endopeptidases;
genetics;
metabolism;
Dactinomycin;
pharmacology;
Down-Regulation;
Genes, Reporter;
Humans;
MicroRNAs;
antagonists & inhibitors;
metabolism;
Microtubule-Associated Proteins;
metabolism;
RNA, Messenger;
metabolism;
Signal Transduction;
drug effects;
Transfection;
Valproic Acid;
administration & dosage;
antagonists & inhibitors;
pharmacology
- From:
Journal of Southern Medical University
2018;38(12):1415-1420
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effect of sodium valproate (VPA) on activation of miR-34c-5p/ATG4B signaling pathway and autophagy in SH-SY5Y cells.
METHODS:Routinely cultured SH-SY5Y cells were treated with VPA at different doses for 24 h, and the changes in the mRNA levels of ATG4B and miR-34c-5p and the protein expression of ATG4B were assessed using qRTPCR and immunoblotting, respectively. The effect of transfection with a plasmid containing ATG4B promoter on the promoter activity of ATG4B in VPA-treated SH-SY5Y cells was assessed using the reporter gene assay. The stability of ATG4B mRNA was analyzed with qPCR in SH-SY5Y cells treated with VPA alone or with VPA combined with the transcription inhibitor actinomycin D. The expression level of miR-34c-5p was detected using qPCR in SH-SY5Y cells treated with VPA alone or with VPA combined with miR-34c-5p mimics or antagonist, and the role of miR-34c-5p in VPA-induced ATG4B down-regulation was evaluated. The changes in the level of autophagy were evaluated by detecting LC3-Ⅱ expression in the cells after treatment with VPA or VPA combined with miR-34c-5p antagonist.
RESULTS:VPA dose-dependently down-regulated the expression of ATG4B at both the mRNA and protein levels in SH-SY5Y cells. VPA treatment did not significantly affect the promoter activity of ATG4B, but obviously lowered the mRNA stability of ATG4B in SH-SY5Y cells. VPA treatment up-regulated the expression of miR-34c-5p, and the miR-34c-5p antagonist reversed VPA-induced down-regulation of ATG4B in SH-SY5Y cells. VPA also down-regulated the expression level of LC3-Ⅱ in SH-SY5Y cells.
CONCLUSIONS:VPA suppresses autophagy in SH-SY5Y cells possibly via activating miR-34c-5p/ATG4B signaling pathway.
