title: [Effects of myoblast determining gene and connexin 43 gene on fibroblast differentiation and biological function in rats].
abstract: OBJECTIVE
  To investigate a change in the differentiation and biological function of the cultured rat fibroblast (FB) transfected by the myoblast determining gene (MyoD) and the connexin 43 (Cx43) gene and to explore the possible mechanism of the MyoD and Cx43 genes on treatment of ischemic heart disease (IHD).
  
  
  METHODS
  The gene cloning technology was used to construct the eukaryotic expressed plasmid vector pLenti6/V5-DEST-MyoD and pLenti6/V5-DEST-Cx43 in which MyoD cDNA or Cx43 cDNA was inserted. The RFL-6 FB cells were transfected with exogenetic MyoD cDNA or Cx43 cDNA via lipofectamine, followed by the Blasticidin (50 microg/ml) selection, according to the lentiviral expression system (ViraPower) protocol. The expression and the biological functions of Myol) and Cx43 in the transfectants were testified by RT-PCR, Western blot, and molecular and immunocytochemical methods. The mophological structure changes of the cells were observed under microscope before and after the transfection.
  
  
  RESULTS
  The expression of MyoD and Cx43 was detected in the MyoD and Cx43 genes transfected FB with RT-PCR and Western blot. The immunocytochemical methods indicated the expressions of the MyoD and Cx43 genes, while desmin and alpha-actin were found in these cells. The myotubes were found from the cultures incubated a week in the differentiation medium, in which the transfected cells had a characteristic of the filaments in their cytoplasm and showed a myoblast morphology.
  
  
  CONCLUSIONS
  MyoD cDNA can induce the cultured FB to differentiate into the myoblasts and Cx43 cDNA can enhance the gap junctional intercellular communication between the cell and the cell. Thus, a further experimental foundation for the therapy of IHD can be provided.
authors: Department of Cardiovascular and Thoracic Surgery, East Hospital Affiliated to Tongji University, Shanghai, China. jinyi_wang@126.com
authors: Jinyi
authors: J
authors: Wang
authors: Huimin
authors: H
authors: Fan
authors: Zhongmin
authors: Z
authors: Liu
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002454:Cell Differentiation; D002478:Cells, Cultured; D018031:Connexin 43 / Q000096:biosynthesis / Q000235:genetics*; D015870:Gene Expression; D015316:Genetic Therapy; D005822:Genetic Vectors; D016086:Lentivirus / Q000235:genetics; D017570:MyoD Protein / Q000096:biosynthesis / Q000235:genetics*; D032446:Myoblasts / Q000166:cytology* / Q000378:metabolism; D017202:Myocardial Ischemia / Q000628:therapy*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
chemical_list: D018031:Connexin 43; D017570:MyoD Protein; D012333:RNA, Messenger