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MiR-200c regulates ROS-induced apoptosis in murine BV-2 cells by targeting FAP-1.
- Source :
- Spinal Cord; Mar2015, Vol. 53 Issue 3, p182-189, 8p
- Publication Year :
- 2015
-
Abstract
- Objective:Reactive oxygen species (ROS) are significantly upregulated after spinal cord injury (SCI). MicroRNAs (miRNAs) are reported to be widely involved in regulating gene expression. This paper aims to explore the correlation between ROS-induced cell apoptosis and abnormal miRNA expression after SCI.Methods:To profile the expression of miRNAs after SCI, miRNA microarray was applied and the result was verified by reverse transcription quantitative PCR (RT-qPCR). ROS production following H<subscript>2</subscript>O<subscript>2</subscript> stimulation was examined using dihydroethidium staining and flow cytometry. The levels of miR-200c after H<subscript>2</subscript>O<subscript>2</subscript> treatment were determined using RT-qPCR. Cell viability and apoptosis were examined in murine BV-2 cells transfected with miR-200c mimics, inhibitor or negative control. Immunofluorescence and western blot were used to further explore the effects of miR-200c on Fas-associated phosphatase-1 (FAP-1) expression.Results:MiR-200c was showed to be significantly increased after SCI by miRNA microassay and RT-qPCR. ROS production enhanced miR-200c expression in a dose-dependent manner and induced significant apoptosis in BV-2 cells. The upregulation of miR-200c reduced cell viability and induced BV-2 cell apoptosis. MiR-200c negatively regulated the expression of FAP-1, thereby inducing FAS signaling-induced apoptosis. RT-qPCR analysis showed that the FAP-1-targeting small interfering RNA (siRNA) did not affect the level of miR-200c in murine BV-2 cells. In addition, suppression of FAP-1 by siRNA promoted apoptosis, even in cells that were co-transfected with the miR-200c inhibitor.Conclusions:The current data suggested that miR-200c contributes to apoptosis in murine BV-2 cells by regulating the expression of FAP-1. This proposes a therapeutic target for enhancing neural cell functional recovery after SCI. [ABSTRACT FROM AUTHOR]
- Subjects :
- REACTIVE oxygen species
ANALYSIS of variance
ANIMAL experimentation
APOPTOSIS
CELL culture
CELL physiology
FLOW cytometry
FLUORESCENT antibody technique
GENE expression
HYDROGEN peroxide
MICE
PHOSPHATASES
POLYMERASE chain reaction
RESEARCH funding
RNA
SPINAL cord injuries
STATISTICS
WESTERN immunoblotting
DATA analysis
OXIDATIVE stress
DATA analysis software
MICROARRAY technology
DESCRIPTIVE statistics
Subjects
Details
- Language :
- English
- ISSN :
- 13624393
- Volume :
- 53
- Issue :
- 3
- Database :
- Complementary Index
- Journal :
- Spinal Cord
- Publication Type :
- Academic Journal
- Accession number :
- 102365004
- Full Text :
- https://doi.org/10.1038/sc.2014.185