Effect of small interfering RNA against connective tissue growth factor in TGF-β_1 stimulated human keloid fibroblasts

Objective To investigate the effect of small interfering RNA(siRNA)-expressing plasmid targeting connective tissue growth factor(CTGF) on the expressions of its mRNA and protein in cultured human keloid fibroblasts(hKFs).Methods Constructed siRNA-expressing plasmids targeting CTGF were transfected into hKFs by Dosper package.Then the transfected cells were treated with transforming growth factor 1(TGF-β1,10 ng/ml).Normal hKFs cells and those with TGF-β1 treatment served as normal control and control groups respectively.The expression level of CTGF mRNA and protein were detected by real-time fluorescent quantitative PCR(RQ-PCR) and Western blotting respectively.Results The expression level of CTGF mRNA was only 56.6% of that of the normal control(P0.05),and the level of CTGF protein was close to that of the normal control(P0.05).Conclusion Our constructed CTGF siRNA plasmid suppresses the mRNA and protein expressions of the target gene in cultured hKFs,even in the cells after TGF-β1 stimulation.This approach may be used as a novel method to treat keloid by gene therapy.