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Proteomic analysis of anti‐angiogenic effects by a combined treatment with vinblastine and rapamycin in an endothelial cell line

Angiogenesis controls the new blood supply routes into the tumor mass via the host endothelial cells (ECs). In this study, the EA.hy926 endothelial cell line has been treated with vinblastine (VBL) and rapamycin (RAP), both separately and in combination at low doses. Recently, we demonstrated the synergistic antiangiogenic effects of a combination of VBL and RAP at very low doses in vitro and in vivo. Herein, we confirm the ability of this combined treatment to statistically inhibit the proliferation of ECs, in a synergistic manner, by inducing apoptosis. The aim of this study was to substantiate these findings at the protein level. Differential proteomic analysis was performed on untreated control cells, treated with VBL, incubated with RAP, or subjected to a drug combination. Differentially expressed 113 polypeptide chains were visualized and 65 were identified via MALDI‐TOF analysis. Some of the regulated proteins are involved in the processes of angiogenesis, proliferation, migration, and apoptosis. The down‐modulation of ATP synthase, annexin A2, heat shock p70, glucose‐6‐phosphate dehydrogenase, vasodilator‐stimulated phosphoprotein, proteasome 26S, tryptophanyl‐tRNA synthetase, and stathmin/OP18, as well as the up‐modulation of carbonyl reductase, Rho‐GDI, and histone H1.0 correlates with the synergistic antiangiogenic activity of VBL and RAP.