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Yi Zhu, Chuan Chen, Ying Lin, Yizhi Liu, Mingxing Wu, Cataract Lab; Downregulation of Syndecan-4 by RNA Interference Inhibits Adhesion and bFGF-induced Proliferation of Lens Epithelial Cells. Invest. Ophthalmol. Vis. Sci. 2013;54(15):479. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
The present study investigates the role of syndecan-4 in adhesion and basic fibroblast growth factor (bFGF)-induced proliferation of lens epithelial cells (LECs).
The expression of syndecan-4 on LECs (SRA 01/04 cell line) was detected by reverse transcription-polymerase chain reaction (RT-PCR). The siRNA sequence targeting syndecan-4 was transfected to LECs. The expression of syndecan-4 after RNA interference (RNAi) was measured by western blot. Cell attachment assay was performed, and cell adhesion rate was observed by inverted microscope and measured by MTT assay. Proliferation of LECs induced by bFGF was measured by MTT assay. Cell-cycle was analyzed by flow cytometry. The expression of proliferating cell nuclear antigen (PCNA) was measured by western blot. The expression of protein kinase Cα(PKCα) was analyzed by RT-PCR.
Syndecan-4 was expressed in SRA 01/04 cells, and the expression was significantly reduced after RNAi. Cell attachment was suppressed by knockdown syndecan-4. The proliferative effect of bFGF on cells was significantly reduced at 48h by syndecan-4 siRNA(p<0.01), with the inhibition rate at 15.7%. Cell-cycle was delayed and the expression of PCNA was suppressed. Also, bFGF-induced expression of PKCα was reduced (p<0.01) by knockdown syndecan-4.
Downregulation of syndecan-4 by RNAi can inhibit adhesion and bFGF-induced proliferation of SRA 01/04 cells. The possible mechanism may be insufficient activation of PKCα. Further study of syndecan signal transduction pathway may provide a new approach for prevention and treatment of capsular opacification.
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