title: MicroRNA-497 Induces Apoptosis and Suppresses Proliferation via the Bcl-2/Bax-Caspase9-Caspase3 Pathway and Cyclin D2 Protein in HUVECs.
abstract: BACKGROUND
  MicroRNAs play crucial roles in various types of diseases. However, to date, no information about the role of miR-497 in the development of atherosclerosis has been reported. This study investigated the possible role of miR-497 in vascular endothelial cell injury during the early stage of atherosclerosis.
  
  
  METHODS
  The expression level of miR-497 in human umbilical vein endothelial cells (HUVECs) exposed to ox-LDL was detected using qRT-PCR. To perform gain of function and loss of function analyses, miR-497 mimics were transfected into HUVECs, and miR-497 inhibitors were transfected into HUVECs stimulated with ox-LDL. Flow cytometry was used to analyze cell cycle progression and apoptosis. EdU and CCK-8 assays were employed to detect DNA synthesis and cell proliferation, respectively. After bioinformatics prediction, a dual Luciferase Reporter assay was used to analyze the direct target genes of miR-497. The mRNA and protein levels of the target genes were detected using qRT-PCR and western blot analyses, respectively. Caspase-9/3 activity was analyzed to determine the mechanism of endothelial dysfunction.
  
  
  RESULTS
  We showed that miR-497 was significantly upregulated in HUVECs stimulated with ox-LDL. Ectopic expression of miR-497 suppressed cell proliferation, induced apoptosis and increased the activity of caspase-9/3. After verification, Bcl2 and CCND2 were shown to be direct target genes of miR-497 in HUVECs. MiR-497 significantly suppressed cell proliferation by arresting the cell cycle through the CCND2 protein and induced apoptosis through the Bcl2/Bax-caspase9-caspase3 pathway.
  
  
  CONCLUSIONS
  Overall, our study shows that miR-497 might play a role in the development of atherosclerosis by inducing apoptosis and suppressing the proliferation of vascular endothelial cells. Therefore, miR-497 could be a potential therapeutic target for the treatment of atherosclerosis.
authors.affiliation: Department of Vascular Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, P. R. China.
authors.forename: Ridong
authors.initials: R
authors.lastname: Wu
authors.affiliation: Department of Breast Surgery, Dongguan Maternal & Children Health Hospital, Dongguan, P. R. China.
authors.forename: Shi
authors.initials: S
authors.lastname: Tang
authors.affiliation: Department of Vascular Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, P. R. China.
authors.forename: Mian
authors.initials: M
authors.lastname: Wang
authors.affiliation: Department of Vascular Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, P. R. China.
authors.forename: Xiangdong
authors.initials: X
authors.lastname: Xu
authors.affiliation: Department of Vascular Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, P. R. China.
authors.forename: Chen
authors.initials: C
authors.lastname: Yao
authors.affiliation: Department of Vascular Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, P. R. China.
authors.forename: Shenming
authors.initials: S
authors.lastname: Wang
mesh_terms: D017209:Apoptosis / Q000235:genetics*; D050197:Atherosclerosis / Q000235:genetics; D053148:Caspase 3 / Q000235:genetics*; D053453:Caspase 9 / Q000235:genetics*; D049109:Cell Proliferation / Q000235:genetics*; D002478:Cells, Cultured; D056742:Cyclin D2 / Q000235:genetics*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D008077:Lipoproteins, LDL / Q000235:genetics; D035683:MicroRNAs / Q000235:genetics*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics*; D012333:RNA, Messenger; D051028:bcl-2-Associated X Protein / Q000235:genetics*
chemical_list: C080336:CCND2 protein, human; D056742:Cyclin D2; D008077:Lipoproteins, LDL; C559640:MIRN497 microRNA, human; D035683:MicroRNAs; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D051028:bcl-2-Associated X Protein; C099252:oxidized low density lipoprotein; D053148:Caspase 3; D053453:Caspase 9