Experimental Study On The Synergistic Inhibitory Effects Of BFGF Monoclonal Antibody Plus S-1 On Lewis Lung Cancer

Objectives:To research the inhibitory effect of the proliferation and the effect of the apoptotic induction on the Lewis lung cancer cell line (LLC) in vitro by bFGF monoclonal antibody MabF7 and S-1 alone or in combination correspondingly. To observe the inhibitory effects of the growth, metastasis, angiogenesis in Lewis lung carcinoma of the mice by bFGF monoclonal antibody MabF7 and S-1 alone or in combination correspondingly. To investigate the synergistic inhibitory effects and the probable mechanism by the experimental study about the inhibition effects of MabF7 plus S-1 on Lewis lung cancer in vivo and vitro.Methods:1. CCK-8 was used to assess the inhibitory effects of bFGF mAb MabF7 and S-1 alone and in combination on Lewis lung cancer cell (LLC) proliferation; The LLC cell apoptosis was analyzed by flow cytometry with Annexin V/PI staining.2. The spontaneous metastatic graft model of twenty-eight C57BL/6 mice bearing Lewis lung cancer were established and when the tumor was touched the mice were divided randomly into four groups:normal sodium(NS) treatment group, bFGF mAb MabF7(MabF7)treatment group, S-1 treatment group, bFGF mAb MabF7 plus S-1 (MabF7+S-1)treatment group. Each group were administered for 15 days, MabF7treatment group was injected MabF75 times,1 time/3 days. Each of the remaining groups were administered for 15 days continuously. Body weight and tumor volume of the mice were measured every three days in thel5days treatment; The change curve of the body weight and the tumor volume were charted; Tumor tissues were isolated on the 18th day, weighted. The inhibitory rate of body weight and tumor growth were calculated and charted;The metastases on the surface of Lewis lung carcinoma were counted. CD31 staining endothelial cell was used to assay microvessel density (MVD). Results:1. Concentrations of 50,100,150 and 200μg/ml of bFGF mAb MabF7 inhibited proliferation of Lewis lung cancer cells in vitro and was in a concentration-dependent manner. Concentrations of 5,10,20μg/ml of S-1 inhibited proliferation of Lewis lung cancer cells and was in a concentration-dependent manner. The combined inhibitory effects of bFGF mAb MabF7 at a concentration of 100μg/ml and S-1 at the gradient concentration of 5,10,20μg/ml on cell proliferation were more significant than that of MabF7 or S-1 group (P<0.005);2. Flow cytometry results showed that MabF7 at a concentration of 100μg/ml and S-1 at a concentration of 10μg/ml could induce the apoptosis of LLC cells; the two had synergistic apoptotic effect.3. MabF7 and S-1 alone and in combination had inhibitory effects on the growth of the tumor, the inhibitory rate of tumor weight of MabF7 group, S-1 group, (MabF7+S-1) group was 37.80%,47.71%,65.93% respectively, it was more significant in the combination group(P<0.05).4. Each group of MabF7, S-1, (MabF7+S-1) had inhibitory effects on the metastasis of lung nodules, the inhibitory rate was 25.81%,46.77%,69.35% respectively, it was more significant in the combination group(P<0.05).5. CD31 staining endothelial cell was used to detect microvessel density (MVD),the results were:the MVD of MabF7 group (33.43±4.86) was significantly lower than NS group (67.00±5.51) (P=0.000); the MVD of S-1 group (41.86±6.31) compared with the NS group was different significantly (P=0.000); the MVD of (MabF7+S-1) group compared with the NS group was also different significantly (P =0.000), while the MVD of (MabF7+S-1) group was significantly lower than the other treated groups (P<0.005).Conclusions:1. Both bFGF mAb MabF7 and S-1 had inhibitory effects on the proliferation of Lewis lung cancer cell in vitro and were in a concentration-dependent manner. The combination of MabF7 and S-1 had synergistic inhibitory effects on the cell proliferation; MabF7 and S-1 could induce apoptosis in Lewis lung cancer cells in vitro. The combination of MabF7 and S-1 had synergistic effects on the induction of apoptosis.2. MabF7 and S-1 could inhibit the growth and migration of the tumor in the mice, the combination of MabF7 and S-1 had synergistic inhibitiory effects on tumor growth and migration. The mechanism was concerned with the inhibitory effects of MabF7 and S-1 on tumor angiogenesis.