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Purpose: To investigate the effect of small interfering RNA (siRNA)targeting VEGF of retinal pigment epithelium (RPE) cells on the growth activity of human retinal vascular endothelial cells (RECs) under a co-culture system.Methods: By applying the vector (pGPU6)-based siRNA plasmid gene silence system, we specifically silenced VEGF expression of RPE cells (ARPE-19) through plasmid (pGPU6-VEGFA-siRNA)transfection. Reverse transcription polymerase chain reaction (RT-PCR) was applied for selecting the most efficient siRNA segment among three pGPU6-VEGF-siRNA groups (siRNA-1, siRNA-2 and siRNA-3). Treated RPE cells were co-cultured with RFCs in a co-culture system made up of a 24-well culture plate and transwell inserts assembled inside During 7-day culture period, the growth capacity of RECs were observed and tested in the form of cell counting assay. Three groups were established in this study:RPE cells transfected with pGPU6-VEGF-siRNA and co-cultured with RFCs (group A), RPE cells transfected with siRNA null vector and co-cultured with RECs (group B), and RECs cultured alone (group C).Results: After transfection, VEGF expression levels of RPE cells in three pGPU6-VEGF-siRNA groups (siRNA-1, siRNA-2 and siRNA-3)evaluated by RT-PCR were 2.56±0.45,1.17±0.38 and 4.39+0.51, respectively (n=10). siRNA-2 was selected as the foremost segment for transfection (P ＜0.05,SNK-q test). During the 7-day co-culture period, the influence upon the growth of RECs was observed. Growth curve of RECs under co-culture showed a lower growth rate in group A than in group B(P＜0.05,dunnett's test), but no significant difference between group A and group C was noted (P＞0.05,dunnett's test). RECs in group A proliferated much faster during the first four days post-transfection.Conclusion: Delivery of siRNA targeting VEGF plays an efficient role in down-regulating VEGF expression in RPE cells,therefore modulating the growth activity of RECs under a coculture system in vitro. The application of this technique may provide novel evidence for the prevention and treatment of retinal neovascularization diseases.