Objective To observe the influence of secreted endostatin (ES) from liver stem cells as gene therapy vector on proliferation and apoptosis of vascular endothelial cells.Methods To establish liver stem cells by recombinant adenovirus, which could express endostatin stably, and were named WB-ES by our department.WB-ES cells were cultured to acquire the supernatant including endostatin.Then, to add the different concentration supernatant into ECV304 cells for further culturing for 24 h, 48 h, 72 h respectively, and equivalent PBS was added in the control group.The inhibitory effect of supernatant was measured by methyl thiazolyl tetrazolium (MTT) assay.Cell cycle analysis of the experiment group and the control group ECV304 cells was detected by flow cytometry, and early apoptosis was detected by Annexin V-FITC immunofluorescence flow cytometry.Results Free endostatin inhibited the proliferation of ECV304 cells in vitro, which was expressed in the supernatant of liver stem cell WB-ES.Inhibitory effect had the features of concentration dependent and reached the summit at 48 h.In the experiment group, endostatin induced the G1 phase block of cell cycle and decrease of S phase cells proportion (P<0.05) .Early apoptosis rate was higher in the experiment group, but there was no statistical difference in the two groups.Conclusion secreted endostatin expressed by WB-ES could inhibit the proliferation of endothelial cells effectively, which illustrated the potential ability of WB-ES as a vector for anti-angiogenesis of HCC gene therapy.
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