Effect of cladribine on growth inhibition and autocrining cytokines in hu-man umbilical vein endothelial cell line EA.hy926 / 中国病理生理杂志

ABSTRACT

AIM: To study the effects of cladribine on growth and secretion activity of human umbilical vein endothelial cell line EA.hy926, and to investigate the mechanism of its anti-tumor effect by inhibiting endothelial cells. METHODS:The effects of cladribine at different concentrations on the cell viability were detected by CCK -8 assay.Apop-tosis and cell cycle distribution were examined by flow cytometry.The protein expression levels were determined by Western blot.The levels of tumor necrosis factor-α(TNF-α), transforming growth factor-β1(TGF-β1)and vascular endothelial growth factor(VEGF)secreted by EA.hy926 cells with cladribine treatment for 48 h were analyzed by ELISA.The nitric oxide(NO)production was measured by Gries method.RESULTS:Cladribine at 0.4~1 μmol/L inhibited the viability of EA.hy926 cells in time-and dose-dependent manners.The IC50was about 3.644 μmol/L.The results showed 43.74% cells in S phase when the concentration of cladribine was 0.4 μmol/L,and 77.23 % cells in S phase when the concentra-tion of cladribine was 1 μmol/L.The apoptosis was not induced by cladribine at 0.4~10 μmol/L.The protein expression of Bax and caspase-3 did not change.The expression of p21 increased and the p53 decreased(P<0.05).The levels of TNF-αand TGF-β1 secreted by EA.hy926 cells increased after cladribine treatment for 48 h.The levels of VEGF and NO decreased.CONCLUSION:Cladribine obviously inhibits the viability of EA.hy926 cells.The mechanism is related to the cell cycle arrest.Cladribine promotes the secretion of TNF-αand TGF-β1 by EA.hy926 cells and inhibits the secretion of VEGF and NO.