BFGF siRNA expression plasmid inhibits growth of human LEC-B3 cells

Abstract

Basic fibroblast growth factor (bFGF) has been thought to play an important role during the development of posterior capsule opacification (PCO) by promoting the growth of lens epithelial cells (LECs). In the present study, we sought to explore the inhibition effect on LEC-B3 cells growth by plasmid-based RNA interference (RNAi) targeting bFGF. It is a prospective study. LEC-B3 cells, an immortal human lens capsule epithelial cell line, were identified as human lens epithelium by immunohistochemical assay of alphaB-crystalline. Both bFGF and bFGF receptor 1 (FGFR1) mRNA expressions of LEC-B3 cells were determined by reverse transcription polymerase chain reaction (RT-PCR). Based on the identification of bFGF expression, plasmid-based RNA interference (RNAi) was used to inhibit bFGF mRNA expression of LEC-B3 cells by stable transfection. bFGF mRNA and protein levels were examined by real time PCR and western blot, respectively. Then the flow cytometry (FCM) was performed to evaluate proliferation cell nuclear antigen (PCNA) expression. PCNA level in bFGF-interfered LEC-B3 cells was compared to that of the control. P < 0.05 was regarded as statistically different. bFGF and FGFR1 mRNAs were abundantly expressed in LEC-B3 cells. bFGF mRNA and protein levels were both down-regulated by stable transfection of bFGF siRNA expression plasmid. PCNA expression was significantly decreased by inhibiting bFGF expression (t = -5.0011, P = 0.0005). bFGF and FGFR1 mRNA are extensively expressed in LEC-B3 cells. Plasmid-based RNAi targeting bFGF can lead to potent inhibition of LEC-B3 cells growth, which may play a part in dealing with the development of PCO.