Histone Deacetylase Inhibitor SAHA Suppresses The Angiogenesis Of HUVEC

ObjectiveTo study the effect of histone deacetylase inhibitor (HDACi) SAHAon the cell cycle and apoptosis of human umbilical vein endothelial cell(HUVEC); To observe the effect of SAHA on the proliferation, migrationand tube formation of HUVEC; To further investigate the effect of SAHAon the angiogenesis in the mice. The relationship and initiative mechanismbetween HDACi SAHA with angiogenesis needs to be explored.Methods1. The proliferation of HUVEC affected by SAHA was investigated byCCK-82. The distribution of cell cycle in HUVEC treated with SAHA for48hours was tested by flow cytometry; the extent of apoptosis was evaluatedby Annexin V-fluorescein isothiocyanate and propidium iodide staining.3. To investigate the effect of SAHA on the migration of HUVEC:transwell assay was performed to observe the migration of HUVEC whichwas treated with15μM SAHA for48hours.4. To evaluate the effect of SAHA on the tube formation of HUVEC in vitro: the ability of tube formation of HUVEC was tested by the matrigelangiogenesis assay in vitro, after the HUVEC was treated by15μM SAHAfor48hours.5. To observe the effect of SAHA on the change of protein expressionin HUVEC: after treated with15μM SAHA for48hours, Western-blot wasused to evaluate the change of caspase, VEGF-induced signaling pathway,P53and P21.6. To observe the effect of SAHA on the tube formation in vivo: thematrigel angiogenesis assay in vivo was operated to observe the newformed vessel in matrigel plug, thereby evaluating the effect of SAHA onthe angiogenesis in mice.Results1. The results of CCK-8assay indicated that SAHA can significantlysuppress HUVEC proliferation in a dose-dependent manner andtime-dependent manner.2. The results of flow cytometry suggested that the rate of S-phase cellwas increased; the results of Annexin V-fluorescein isothiocyanate andpropidium iodide staining imply that the apoptosis of HUVEC wasremarkably enlarged.3. The results of transwell assay demonstrated that the number ofmigrated HUVEC was significantly decreased after the treatment of15μMSAHA for48hours. 4. The matrigel angiogenesis in vitro suggested that the capillary-liketube formation in the SAHA treated group was less than the control group.5. Western-blot results reflected that SAHA increased the volume ofactive casapase and inhibited the VEGF-induced signaling pathway, at thesame time, SAHA promoted the expression of P21and P53.6. The matrigel angiogenesis in vivo indicated that the number of newformed vessels in the matrigel plug in SAHA treated group was less than inthe control group.ConclusionThe proliferation, migration and angiogenesis in vitro of HUVECwere inhibited by the HDACi SAHA. SAHA can arrest the cell cycle ofHUVEC in S-phase, and SAHA can promote the apoptosis of HUVEC. Thepotential mechanism, SAHA inhibits HUVEC in vitro and suppresses theangiogenesis in vivo, may involve the SAHA block of the VEGF-inducedsignaling pathway and trigger of caspsase cascade, meanwhile, with theexpression of P53and P21.