The Anti-Tumor Immunity Effect Of Dendritic Cells Vaccine Loaded With CTP-FoxM1 On Mice Implanted Subcutaneous Tumor Model

Objective: To investigate the expression of FoxM1 antigen in the mouse Hepatoceellular carcinoma cell line Hepa1-6, and evaluate its possibility to establish subcutaneous tumor model. To explore the optimum condition that established of mouse subcutaneous hepatocellular carcinoma model with Hepa 1-6 cells. To research the anti-tumor immune effects of CTP-FoxM1 loaded DCs vaccine on the mice hepatocellular carcinoma.Methods: Hepa 1-6 cells were cultured in DMEM media containing10% fetal calf serum and 1% penicillin-streptomycin( 37℃、5%CO2),the FoxM1 protein level of Hepa 1-6 HCC cell Line was analyzed with western blot. The C57/BL6 mice were inoculated subcutaneously with o.1ml Hepa 1-6 cell suspension at the concentration of 5x106/ml 、1x107/ml、3x107/ml, to confirm the optimum condition of establish tumor model. CTP-FoxM1 fusion protein was incubated with mice bone marrow dendritic cells(BMDC) for 48 h as the DC vaccine. Hepa 1-6 cells were injected subcutaneously into the C57BL/6 mice, 7 days after the inoculation, DC vaccine was injected to subcutaneous of mice, tumorvolumes were monitored twice day. When the animal experiments were finished, the tumor mass were isolated and weighed, the tumor growth curve was draw, the serum level of TNF-α、IFN-γ and IL-2 were measured by ELISA. Lung, liver and spleen were process pathological examination with hematoxylin-eosin(HE) staining to assess the tumor metastasis. In addition, the levels of serum CXCL13 of 78 patients with HCC were measured by enzyme-linked immunosorbent assays. Clinical characteristics, laboratory parameters were analyzed retrospectively and the role of CXCL13 in hepatocellular carcinoma patients was evaluated.Results: High level FoxM1 were expressed in Hepa 1-6 cells. The established of mice subcutaneous tumor with 0.1ml Hepa 1-6 cells at the concentration of 3x107/ml has a high success rate, it spend a short time and many mitotic figure could be see. Compared with the other groups,CTP-FoxM1-DC vaccine treated groups showed significantly lower weight and volume of tumor mass. The levels of IFN-γ or IL-2 have no significant difference when CTP-FoxM1-DC group comared with other groups. HE results show that there was apparent inflammation and injury in small intestines which has a low expression of FoxM1. Serum CXCL13 was significantly higher in HCC patients compared with healthy controls and was relative with the tumor size, the metastasis and the TMN stages.Conclusion: The results showed that high level FoxM1 expression in the mice HCC cell line Hepa1-6, it could be used to bulid subcutaneoustumor model which was positive express of FoxM1. Hepa 1-6 cells could be used to build ideal mice subcutaneous tumor model at the concentration of 3x107/ml, laying the foundation for observed the anti- tumor immunity of DC vaccine. Injected with CTP-FoxM1 antigen loaded DC vaccine in tumor bearing mice, more effective anti-tumor immune reactions could be induce in mice than that with other vaccines. The results suggest that the CTP-FoxM1 loaded DC tumor vaccine can prevent development of liver cancer. The CTP-Fox M1 Pulsed DC vaccine are expected to develop a new method for HCC immune therapy. CXCL13 may have clinical values of diagnosis and prognosis in HCC.