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-?1 (TGF-?1) for up to 48 hours. The mRNA levels of Hsp47 and ? smooth muscle actin (?SMA) 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 ?SMA proteins was determined by western immunoblotting. RESULTS: TGF-?1 increased the mRNA expressions of both Hsp47 and ?SMA in human Tenon's fibroblasts, as determined by quantitative real time RT-PCR. However, it induced the protein expression of only ?SMA but not Hsp47, as determined by western immunoblots. When siRNAs specific for Hsp47 were introduced into those cells, the TGF-?1-induced expression of ?SMA was significantly attenuated on western immunoblots; after 48 hours of exposure to TGF-?1, the relative densities of immunobands were 11.58 for the TGF-?1 only group and 2.75 for the siRNA treatment group, compared with the no treatment control group (p?

SUBMITTER: Hong S 

PROVIDER: S-EPMC3490793 | biostudies-other | 2012

REPOSITORIES: biostudies-other