title: Role of heat shock protein 47 in transdifferentiation of human tenon's fibroblasts to myofibroblasts.
abstract: BACKGROUND
  Heat shock protein 47 (Hsp47) is a well-known molecular chaperone in collagen synthesis and maturation. The aim of this study is to investigate its putative role in the transdifferentiation of Tenon's fibroblasts to myofibroblasts.
  
  
  METHODS
  Primary cultured human Tenon's fibroblasts were exposed to transforming growth factor-β1 (TGF-β1) for up to 48 hours. The mRNA levels of Hsp47 and α smooth muscle actin (αSMA) were determined by quantitative real time RT-PCR. After delivery of small interfering RNA (siRNA) molecules targeting Hsp47 into the cells, the expression of Hsp47 and αSMA proteins was determined by western immunoblotting.
  
  
  RESULTS
  TGF-β1 increased the mRNA expressions of both Hsp47 and αSMA in human Tenon's fibroblasts, as determined by quantitative real time RT-PCR. However, it induced the protein expression of only αSMA but not Hsp47, as determined by western immunoblots. When siRNAs specific for Hsp47 were introduced into those cells, the TGF-β1-induced expression of αSMA was significantly attenuated on western immunoblots; after 48 hours of exposure to TGF-β1, the relative densities of immunobands were 11.58 for the TGF-β1 only group and 2.75 for the siRNA treatment group, compared with the no treatment control group (p < 0.001).
  
  
  CONCLUSIONS
  Our data suggest that Hsp47 may be related to the TGF-β1-induced transdifferentiation of human Tenon's fibroblasts to myofibroblasts.
authors.affiliation: Institute of Vision Research, Department of Ophthalmology, Yonsei University College of Medicine, 50 Yonsei-ro, Seodaemun-gu, Seoul, Republic of Korea.
authors.forename: Samin
authors.initials: S
authors.lastname: Hong
authors.forename: Kyoungsoo
authors.initials: K
authors.lastname: Park
authors.forename: Jin Hyoung
authors.initials: JH
authors.lastname: Kim
authors.forename: Sueng-Han
authors.initials: SH
authors.lastname: Han
authors.forename: Jong Bok
authors.initials: JB
authors.lastname: Lee
authors.forename: Gong Je
authors.initials: GJ
authors.lastname: Seong
mesh_terms: D015153:Blotting, Western; D054338:Cell Transdifferentiation / Q000502:physiology*; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D005786:Gene Expression Regulation; D050958:HSP47 Heat-Shock Proteins / Q000502:physiology*; D006801:Humans; D058628:Myofibroblasts / Q000166:cytology* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D050958:HSP47 Heat-Shock Proteins; D012333:RNA, Messenger