Effects of RNA interference on CTGF expression and collagen metabolism of keloid fibroblasts

Abstract

Objective To investigate the influence of connective tissue growth factor (CTGF) onthe collagen metabolism in human keloid fibroblasts with RNA interference (RNAi).Methods Human keloid fibroblasts (KFB) in vitro were transfected by 3 pairs of specific small interfering RNA (siRNA) CTGF plasmid synthesized for human CTGF,respectively.Reverse transcriptase-polymerase chain reaction (RT-PCR) contributed to the screening of the best siRNA in interfering of CTGF expression in human keloid fibroblasts to construct the plasmids,with the application of RNAi,to test the changes of expression level and collagen content of CTGF in transfected keloid fibroblasts through RT-PCR and Western blotting compared to its control groups.Results The 3rd pair (C3) siRNA- CTGF expression of genes and proteins was remarkably inhibited after being interfered with human keloid fibroblasts,with inhibitory rates of 86.8 ％ and 65.6 ％.Conclusions Keloid fibroblasts transfected by plasmid siRNA-CTGF effectively inhibite the expression of CTGF and deposition of collagen,and CTGF promotes the collagen synthesis in keloid development. Key words: Keloid; Fibroblasts; RNA interference; Connective tissue growth factor