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Inhibitory effect of rapamycin on the proliferation of Rhesus retinal vascular endothelial cells

Background The signal pathway of mammalian target of rapamycin (mTOR) plays an important role in the regulation of cell cycle.Rapamycin(RAPA) can inhibit the proliferation of cells by regulating of cell cycle.Objective This study was to investigate the effect of RAPA on the proliferation of Rhesus retinal vascular endothelial cells (RF/6A).Methods RF/6A cell strains were cultured in vitro,and PBS,10 μg/L RAPA or 5 μg/L RAPA was added into the medium respectively.The expression of cyclin D1 protein in the RF/6A cells (absorbancy) were detected by Western blot.The cell cycle distribution after RAPA action was analyzed by flow cytometry.Matrigel was used in endothelial-cell tube formation to evaluate the effect of RAPA on angiogenesis.Results Western blot assay showed that the expressions of cyclin D z protein were 0.92±0.04,0.58±0.02 and 0.73±0.02 in the PBS group,10 μg/L RAPA group and 5 μg/L RAPA group,showing a significant difference among the three groups (F =246.320,P =0.000),and the relative expressing level of cyclin D1 protein in the l0 μg/L RAPA group and 5 μg/L RAPA group was significantly lower than that of the PBS group (both at P＜0.05).The proportion of G0/G1 cells were (42.13±0.57)％,(65.15±0.64)％ and (54.09± 0.78)％ respectively in the PBS group,10 μg/L RAPA group and 5 μg/L RAPA group,which was significantly different among the three groups (F=887.815,P=0.000).The number of endothelial-cell tubes were (9.67 ± 1.53)/field,(4.33 ± 0.58)/field and (6.33 ±0.58)/field respectively in the PBS group,10 μg/L RAPA group and 5 μg/L RAPA group,and the number of endothelial-cell tubes in 10 μg/L RAPA group and 5 μg/L RAPA group was less than that in the PBS group,with a significant difference among the three groups (F =21.778,P =0.002).Conclusions RAPA can inhibit the proliferation of RF/6A cells in vitro by down-regulating the expression of cyclin D1. 
 
Key words: 
Vascular endothelial cell;  Retina;  Angiogenesis;  Rapamycin;  Cyclin D1