Abstract:AIM:To observe the effect of platelet-derived growth factor receptor-α antisense oligonucleotide(PDGFR-α ASODN)on the proliferation of rabbit lens epithelial cells N/N1003A in vitro and investigate its mechanism.
METHODS: PDGFR-α ASODN was transfected into N/N1003A cells with LipofectamineTM 2000. The proliferations of the transfected cells were analyzed by MTT assay and the expressions of PDGFR-α mRNA were detected by RT-PCR. Phase microscope and transmission electron microscope were used to observe the changes of cell morphology and ultramicrostructure. Flow cytometry was applied to detect the changes of cell cycle and apoptosis rate.
RESULTS: After N/N1003A cells were treated by PDGFR-α ASODN, the proliferations of the cells were significantly inhibited(P<0.05), and the expression levels of PDGFR-α mRNA were notablely lowered(P<0.05)in a dose-dependent manner, respectively. The cells in the experimental group showed typical features of apoptosis under the transmission electron microscope and the rates of cell apoptosis were markedly higher(P<0.05). The analysis of cell cycle indicated that the cells were arrested in G1 phase.
CONCLUSION: PDGFR-α ASODN can silence PDGFR-α gene expression in rabbit lens epithelial cells, inhibit cell proliferation, and induce cell apoptosis, which provides the experimental basis for the application of PDGFR-α ASODN to prevent the formation of posterior capsular opacification.