1. MiR-200c regulates ROS-induced apoptosis in murine BV-2 cells by targeting FAP-1.
- Author
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Yu, D S, Lv, G, Mei, X F, Cao, Y, Wang, Y F, Wang, Y S, and Bi, Y L
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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 - 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 H2O2 stimulation was examined using dihydroethidium staining and flow cytometry. The levels of miR-200c after H2O2 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]
- Published
- 2015
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