The Effect Of VBP3 Tumor Vaccine In Human Umbilical Vein Endothelial Cells, Melanoma Cells And Tumor-bearing Mice Survival

Object: To study the effect of VBP3 tumor vaccine in human umbilical vein endothelial cells(HUVECs), tumor cell and the survival time of the tumor-bearing mice.Methods: The BALB/c mice were immunized with the purified VBP3 protein, and then prepared the polyclonal antibodies against the VBP3. The antibody titer was detected by ELISA; The inhibitive effects of anti-VBP3 antibodies on HUVECs, B16F0 cells proliferation were detected by CCK-8 in vitro;Transwell assay and the wound-healing assay were used to detect the anti-VBP3 antibodies influence on the migration of B16F0 and HUVECs; Western blot was used to analyze the phosphorylation of Erk1/2 and Akt in the b FGF and VEGF signaling pathways; The survival experiment was to investigate the survival prolongation effect of the VBP3 peptide immunization in the Lewis lung carcinoma transplantable tumor model; the immunohistochemistry was to investigate the microvascular density of the tumor tissue.Result: The result of ELISA showed that high titer anti-VEGF and anti-b FGF antibodies were produced in immunized BALB/c mice; the result of CCK-8 show that the anti-VBP3 antibodies can inhibit the HUVECs and B16F0 cells proliferation in vitro; The results of wound-healing assay showed that the anti-VBP3 antibodies can inhibit the migration of HUVECs and B16F0 cells in vitro; the western-blot results showed that in vitro the anti-VBP3 antibodies inhibited the phosphorylation of Erk1/2 and Akt in HUVECs and B16F0 cells and it had a concentration relationship; the result of the survival experiment showed that the VBP3 can significantly prolong survival of tumor-bearing mice compare to the control group in vivo and inhibite the tumor angiogenesis.Conclusion: The mice immunized with the VBP3 tumor vaccine can generate a high titers of antibodies against the VEGF and b FGF; the anti-VBP3 antibodies can inhibit the B16F0 and HUVECs proliferation and migration in vitro. The VBP3 tumor vaccine also can prolong Lewis lung carcinoma-bearing C57BL/6 mice life time and inhibite the tumor angiogenesis in vivo.