title: MiR-200c regulates ROS-induced apoptosis in murine BV-2 cells by targeting FAP-1.
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 H2O2stimulation was examined using dihydroethidium staining and flow cytometry. The levels of miR-200c after H2O2treatment 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.
authors.affiliation: Department of Orthopaedics, The First Affiliated Hospital, Liaoning Medical University, Jinzhou, People's Republic of China.
authors.forename: D S
authors.initials: DS
authors.lastname: Yu
authors.affiliation: Department of Orthopaedics, The First Affiliated Hospital, Liaoning Medical University, Jinzhou, People's Republic of China.
authors.forename: G
authors.initials: G
authors.lastname: Lv
authors.affiliation: Department of Orthopaedics, The First Affiliated Hospital, Liaoning Medical University, Jinzhou, People's Republic of China.
authors.forename: X F
authors.initials: XF
authors.lastname: Mei
authors.affiliation: Department of Orthopaedics, The First Affiliated Hospital, Liaoning Medical University, Jinzhou, People's Republic of China.
authors.forename: Y
authors.initials: Y
authors.lastname: Cao
authors.affiliation: Department of Orthopaedics, The First Affiliated Hospital, China Medical University, Shenyang, People's Republic of China.
authors.forename: Y F
authors.initials: YF
authors.lastname: Wang
authors.affiliation: Department of Orthopaedics, The First Affiliated Hospital, Liaoning Medical University, Jinzhou, People's Republic of China.
authors.forename: Y S
authors.initials: YS
authors.lastname: Wang
authors.affiliation: Department of Orthopaedics, The First Affiliated Hospital, Liaoning Medical University, Jinzhou, People's Republic of China.
authors.forename: Y L
authors.initials: YL
authors.lastname: Bi