title: [Effect of silencing myocardin gene expression on differentiation of mouse bone mesenchymal stem cells into smooth muscle-like cells induced by PDGF-BB].
abstract: OBJECTIVE
  Construction of a small interfering RNA (siRNA) eukaryotic expression vector specific to mouse myocardin gene and study on the role of myocardin-siRNA on differentiation of mouse bone mesenchymal stem cells (MSCs) into smooth muscle-like cells induced by PDGF-BB in vitro.
  
  
  METHODS
  Mouse MSCs were isolated from bone marrow and cultured with 50 mg/L PDGF-BB and fetal bovine serum (20%). Specific myocardin-siRNA sequence was cloned into a plasmid pGenesil-1.0 vector, which contained U6 promoter. The recombinant plasmid and control plasmid were transfected into MSCs which had been cultured with PDGF-BB for 6 days beforehand. The expression of myocardin mRNA was detected by RT-PCR 48 hours after the transfection. Immunohistochemistry was used to detect the SM-MHC and to identify the smooth muscle-like cells.
  
  
  RESULTS
  The recombinant plasmids carrying myocardin-siRNA sequences were constructed successfully and the myocardin mRNA was reduced 42.86% by pGen-myo-shRNA in comparing with that of the controls (P<0.01); and the expression of SM-MHC protein was down-regulated (P<0.01).
  
  
  CONCLUSIONS
  Subset of mouse MSCs have the potential to differentiate into smooth muscle-like cells, a possible cell source responsible for atherosclerotic plaque formation, and myocardin expression may play an important role during this process.
authors: Institute of Pathology, Affiliated Tongji Hospital, Tongji Medical College, Huazhong Science and Technology University, Wuhan 430030, China.
authors: Guan
authors: G
authors: Huang
authors: Mei
authors: M
authors: Xu
authors: Jun
authors: J
authors: Yu
authors: Han
authors: H
authors: Meng
authors: Xue
authors: X
authors: Chen
authors: Yan
authors: Y
authors: Li
authors: Qiu-rong
authors: QR
authors: Ruan
mesh_terms: D000818:Animals; D000077214:Becaplermin; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D015536:Down-Regulation; D020868:Gene Silencing; D005822:Genetic Vectors; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D009131:Muscle, Smooth, Vascular / Q000166:cytology; D032389:Myocytes, Smooth Muscle / Q000166:cytology* / Q000378:metabolism; D018995:Myosin Heavy Chains / Q000378:metabolism; D009687:Nuclear Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D058226:Plaque, Atherosclerotic / Q000473:pathology; D010957:Plasmids; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D020574:Proto-Oncogene Proteins c-sis; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering; D015534:Trans-Activators / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D014162:Transfection; D000099251:Myocardin
chemical_list: D000077214:Becaplermin; D018995:Myosin Heavy Chains; D009687:Nuclear Proteins; D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D015534:Trans-Activators; D000099251:Myocardin