title: MicroRNA-495 regulates the proliferation and apoptosis of human umbilical vein endothelial cells by targeting chemokine CCL2.
abstract: BACKGROUND
  Endothelium dysfunction plays a critical role in atherosclerosis. MicroRNAs are endogenous non-coding RNAs that suppress gene expression by binding to the 3' untranslated regions of target genes. MiR-495 can regulate the proliferation and apoptosis of cancer cells, however, the roles of miR-495 in endothelial cells (ECs) remain unclear. Therefore, this study aims to investigate the roles and mechanisms of miR-495 on ECs proliferation and apoptosis.
  
  
  METHODS
  MiR-495 and CCL2 expressions were examined using quantitative RT-PCR, ELISA assay and western blot. Bioinformatics analysis and luciferase reporter assay were used to examine the regulatory relationship between miR-495 and CCL2. CCK8 assay, BrdU incorporation assay and flow cytometry were used to analyze the roles of miR-495 and CCL2 on the proliferation of human umbilical vein endothelial cells (HUVECs). The effects of miR-495 and CCL2 on HUVECs apoptosis were examined by tunnel staining and western blot.
  
  
  RESULTS
  MiR-495 was down-regulated in patients with coronary artery disease compared with healthy controls. CCL2 was a novel target gene of miR-495. MiR-495 significantly promoted HUVECs proliferation by altering cell cycle distribution, and it also inhibited HUVECs apoptosis by affecting the expression of cleaved caspase 3. Effects of miR-495 on HUVECs proliferation and apoptosis were significantly reversed by overexpression of CCL2.
  
  
  CONCLUSIONS
  MiR-495 could affect HUVECs proliferation and apoptosis by directly targeting CCL2. This is the first report to disclose the roles and mechanisms of miR-495 on HUVECs proliferation and apoptosis, which may provide a theoretical basis for clarifying the mechanisms of atherosclerosis.
authors.affiliation: Graduate School of Third Military Medical University, Chongqing 400038, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Liu
authors.affiliation: Cardiovascular Research Institute and Department of Cardiology, Shenyang Northern Hospital, Shenyang, Liaoning 110016, China.
authors.forename: Xiao-Lin
authors.initials: XL
authors.lastname: Zhang
authors.affiliation: Cardiovascular Research Institute and Department of Cardiology, Shenyang Northern Hospital, Shenyang, Liaoning 110016, China.
authors.forename: Cheng-Hui
authors.initials: CH
authors.lastname: Yan
authors.affiliation: Cardiovascular Research Institute and Department of Cardiology, Shenyang Northern Hospital, Shenyang, Liaoning 110016, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Li
authors.affiliation: Cardiovascular Research Institute and Department of Cardiology, Shenyang Northern Hospital, Shenyang, Liaoning 110016, China.
authors.forename: Xiao-Xiang
authors.initials: XX
authors.lastname: Tian
authors.affiliation: Cardiovascular Research Institute and Department of Cardiology, Shenyang Northern Hospital, Shenyang, Liaoning 110016, China.
authors.forename: Nan
authors.initials: N
authors.lastname: Zhu
authors.affiliation: Graduate School of Third Military Medical University, Chongqing 400038, China.
authors.forename: Jing-Jing
authors.initials: JJ
authors.lastname: Rong
authors.affiliation: Cardiovascular Research Institute and Department of Cardiology, Shenyang Northern Hospital, Shenyang, Liaoning 110016, China.
authors.forename: Cheng-Fen
authors.initials: CF
authors.lastname: Peng
authors.affiliation: Cardiovascular Research Institute and Department of Cardiology, Shenyang Northern Hospital, Shenyang, Liaoning 110016, China. Electronic address: yalinghan@gmail.com.
authors.forename: Ya-Ling
authors.initials: YL
authors.lastname: Han
mesh_terms: D020413:3' Untranslated Regions; D000368:Aged; D017209:Apoptosis; D050197:Atherosclerosis / Q000097:blood* / Q000276:immunology; D016022:Case-Control Studies; D049109:Cell Proliferation; D018932:Chemokine CCL2 / Q000378:metabolism*; D003324:Coronary Artery Disease / Q000097:blood* / Q000276:immunology; D015536:Down-Regulation; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000378:metabolism*; D008875:Middle Aged
chemical_list: D020413:3' Untranslated Regions; C111835:CCL2 protein, human; D018932:Chemokine CCL2; C577768:MIRN495 microRNA, human; D035683:MicroRNAs
keywords: Apoptosis; Atherosclerosis; CCL2; Endothelial cell; Proliferation; miR-495