title: Involvement of MicroRNA-133a in the Development of Arteriosclerosis Obliterans of the Lower Extremities via RhoA Targeting.
abstract: OBJECTIVE
  RhoA is a critical factor in regulating the proliferation and migration of arterial smooth muscle cells (ASMCs) in patients with arteriosclerosis obliterans (ASO). RhoA is modulated by microRNA-133a (miR-133a) in cardiac myocytes and bronchial smooth muscle cells. However, the relationship between miR-133a and RhoA with respect to the onset of ASO in the lower extremities is uncertain.
  
  
  METHODS
  We employed in situ hybridization (ISH) and immunohistochemistry (IHC) to detect the location of miR-133a and RhoA in ASO clinical samples, respectively. 5-ethynyl-2'-deoxyuridine (EdU), cell counting kit-8 (CCK-8), Transwell and wound closure assays were utilized to determine the features of human ASMC (HASMC) proliferation and migration. The expression of miR-133a in the HASMCs was assessed using quantitative real-time PCR (qRT-PCR), while that of RhoA was examined via qRT-PCR and Western blotting.
  
  
  RESULTS
  We found miR-133a and RhoA to be primarily located in the ASMCs of ASO. miR-133a was significantly downregulated in the ASO tissues and proliferating HASMCs. In contrast, RhoA was upregulated in the ASO samples. The proliferation and migration of HASMCs was markedly promoted by the downregulation of miR-133a and inhibited by the upregulation of miR-133a. The Luciferase assay confirmed that RhoA was a direct target of miR-133a. The upregulation of miR-133a in the HASMCs decreased the RhoA expression at the protein level. Inversely, the downregulation of miR-133a increased the RhoA protein expression. Of note, the overexpression of RhoA in the HASMCs attenuated the anti-proliferative and anti-migratory effects of miR-133a.
  
  
  CONCLUSIONS
  Our data indicate that miR-133a regulates the functions of HASMCs by targeting RhoA and may be involved in the pathogenesis of ASO. These findings may lead to the development of potential therapeutic targets for ASO of the lower extremities.
authors.affiliation: Laboratory of General Surgery, The First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, People's Republic of China; Department of Vascular Surgery, The Affiliated Hospital of Qingdao University, Qingdao, People's Republic of China.
authors.forename: Yongxin
authors.initials: Y
authors.lastname: Li
authors.forename: Mao
authors.initials: M
authors.lastname: Ouyang
authors.forename: Zhen
authors.initials: Z
authors.lastname: Shan
authors.forename: Jieyi
authors.initials: J
authors.lastname: Ma
authors.forename: Jie
authors.initials: J
authors.lastname: Li
authors.forename: Chen
authors.initials: C
authors.lastname: Yao
authors.forename: Zhengrong
authors.initials: Z
authors.lastname: Zhu
authors.forename: Longjuan
authors.initials: L
authors.lastname: Zhang
authors.forename: Lianzhou
authors.initials: L
authors.lastname: Chen
authors.forename: Guangqi
authors.initials: G
authors.lastname: Chang
authors.forename: Shenming
authors.initials: S
authors.lastname: Wang
authors.forename: Wenjian
authors.initials: W
authors.lastname: Wang
mesh_terms: D001162:Arteriosclerosis Obliterans / Q000209:etiology* / Q000378:metabolism* / Q000473:pathology; D015153:Blotting, Western; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D017931:DNA Primers / Q000737:chemistry / Q000235:genetics; D006801:Humans; D007124:Immunoenzyme Techniques; D017403:In Situ Hybridization; D035002:Lower Extremity / Q000503:physiopathology*; D035683:MicroRNAs / Q000235:genetics*; D009131:Muscle, Smooth, Vascular / Q000378:metabolism* / Q000473:pathology; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020742:rhoA GTP-Binding Protein / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
chemical_list: D017931:DNA Primers; C548234:MIRN133 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; C066737:RHOA protein, human; D020742:rhoA GTP-Binding Protein