Transfection of CTGF siRNA inhibits transdifferentiation in human lens epithelium cell line B3 <i>in vitro</i>

Abstract

AIM: To investigate the expression of connective tissue growth factor(CTGF)and α-SMA in human lens epithelium cell(HLEC)line B3 after transfection by liposome-coated siRNA targeting CTGF. METHODS: HLECs were transfected with small interfering RNA(siRNA)targeting CTGF, labeled with 5'- fluorescein isothiocyanate(5'-FITC)and coated with lipofectamine. The transfection ratio was evaluated via fluorescence intensity. Cell counting kit-8(CCK-8)assay was performed to assess cytoviability of both non-transfected and transfected HLECs. Quantitative RT- PCR, cell immunochemistry and Western blot analysis were conducted to detect the expression changes of CTGF and α-SMA after transfection. RESULTS: A highly effective transfection ratio was observed in siRNA co-transfected with lipofectamine. The transfection ratio reached 95% at 24h. The proliferation of HLECs was inhibited by siRNA after 72h transfection. The expression of CTGF and α-SMA significantly decreased in HLECs after transfected by CTGF siRNA for 24h. This effect was not found in negative control siRNA. CONCLUSIONS: SiRNA targeting CTGF decreased CTGF and α-SMA expression in HLECs, which is a potential therapeutic strategy for posterior capsular opacification.

Keywords

• connective tissue growth factor; small interfering RNA; lipofectamine; transdifferentiation
• connective tissue growth factor
• small interfering RNA
• lipofectamine
• transdifferentiation