Synergistic Anti-tumor Activities Of Anti-bFGF Monoclonal Antibodies And Chemotherapy In Vivo And In Vitro

OBJECTIVE:There are many kinds of tumors, like melanomas, lung cancer hepatoma can high level express bFGF and FGFR which closely related to tumor metastasis,staging and prognosis. And bFGF can stimulate the tumor angiogenesis and proliferation of tumor cells. Which suggested that bFGF could be a new target of anti-tumor therapy. Anti-bFGF mAbs developed in our lab have showed good activities in inhibiting proliferation of tumor cells and tumor growth. Here, we mainly discuss the synergy of Anti-bFGF mAbs and chemotherapeutic drugs in vivo and in vitro.METHODS:Melanoma cells were incubated with anti-bFGF mAbs combined with various chemotherapeutic agents, cisplatin,5-Fu and paclitaxel. Cells proliferation assay was carry out on CCK 8 kit. Melanoma cells were subcutaneously inoculated in the flank of C57 BL/6 mice to establish tumor graft mice.The anti-bFGF mAbs were injected with different dosage through caudal vein and subcutaneously (around tumors).The intratumoral microvessel density was analyzed by anti-CD31 antibody staining.Various cell growth factors, like bFGF, bFGFR, VEGF, VEGFR, PDGF, PDGFR were detected to analyze relations and the mechanism of anti-bFGF mAbs. The Leukocytes were isolated by percoll from the mice peripheral blood, spleen and tumor tissue. The expression of CDs was analyzed with different drugs treatment by flow cytometry. The leukocytes from the mice tumor tissues were marked with annexin-V/PI staining and analyzed with flow cytometry. Western blot was used to preliminary analyze the influence of the signal pathway in treated and untreated B16 cells.RESULTS:Melanoma cells were incubated with anti-bFGF mAbs combined with chemotherapeutic agents cisplatin,5-Fu,paclitaxel. The proliferation inhibition rate of melanoma cells in the groups of MabF7, cisplatin,5-Fu, paclitaxe, MabF7+ cisplatin, MabF7+5-Fu, MabF7+paclitaxel, DAM,MabF7+DAM were 10.20-23.2% respectively. After five injections, the tumor inhibiton rate in the groups of blank, MabF7, cisplatin,5-Fu, paclitaxe, MabF7+cisplatin, MabF7+paclitaxel were 94.89% respectively. The microvessel density (MVD) in tumors measured by imunohistochemistry staining (IHCS) showed that the MVD in the groups of PBS, MabF7, DDP, MabF7+DDP, Docetaxel, MabF7+Docetaxel were low respectively. The immunohistochemistry results showed that cell growth factors, like VEGF, FGF-2, FGFR1, PDGFwere low expressed in the groups of Mab and chemotherapeutic drugs. Flow cytometry assays showed that bFGF mAbs have certain effects on the immune escape mechanism. The analysis results of Annexin V/PI staining and western blot showed that bFGF mAbs can induce the apoptosis of tumor cells obviously.CONCLUSIONS:Anti-bFGF mAbs synergistic with chemotherapeutic drugs can effectively inhibit melanoma growth in vivo and in vitro. The synergistic function of the antibody and chemotherapy may conducted by degrading angiogenesis and preventing immune escape, which may provide a more effective method in cancer therapy.