Antitumor Effect Of Endostar Combined With Mut1 Antigen Specific DC-T Cells On Lewis Lung Carcinoma In Mice

ObjectiveThe occurrence,invasion and metastasis of tumor are closely related to tumor angiogenesis. The inhibition of angiogenesis is a new hotspot of tumor treatment strategy. Endostar is a drug to inhibit the antitumor activity of vascular growth. Studies showed that monotherapy with antiangiogenic agents is failing to produce longlasting clinical responses in most patients, however, antiangiogenic molecules may contribute to persistent tumor’s remission in combination with immunotherapy. And it suggested that antiangiogenic agents synergized with cellular immunotherapy. Dendritic cells are the most potent antigen-presenting cells, playing an important role in the regulation of innate and adaptive immune response.Dendritic cells loaded with special tumor antigen are one of the biological treatment in recent years.In this article, we explored the relationship between Endostar and cellular immunotherapy on antitumor treatment of lewis lung carcinoma in mice, the role and molecular mechanisms of Endostar playing its antitumor effect in the tumor microenvironment, then provided a theoretical basis for the main targets and mechanism of Endostar antitumor effect and promoting the longlasting and potent therapeutic efficacy of Endostar combined with cellular immunotherapy. we may provide a theoretical basis for the optimal strategy of raising the longlasting and potent therapeutic efficacy of Endostar combined with cellular immunotherapy.Methods1.The culture of Lewis lung cancer cells (LLC):Observed the state and the number of the cells in time.The medium was changed and passaged in a timely manner.When the cells went into the logarithmic growth phase,it was prepared for the experiment.2. The culture of mutl antigen loaded DC-T cells:Isolation and culture of mouse spleen T lymphocytes, isolation and culture of mouse dendritic cells, preparation of mutl antigen specific DC-T cells co cultured with T lymphocytes to mutl antigen loaded DC-T cells.3.The transplanted Lewis lung cancer models of C57BL/6 mice were established by left extremity axillary subcutaneous injection of Lewis lung cancer cells (LLC). The tumor-bearing mice were randomly divided into three groups, including A group(PBS control group), B group(DC-T group), and C group(DC-T+Endostar group).4.The situation of tumor growth and the measure of tumor suppression rate:The body weight and tumor diameter were measured every other day, respectively.24 hours after the last treatment,mice were sacrificed. Tumor tissue removed and weighed.5. The microvessel density (MVD) in the tumor tissue of tumor-bearing mice was detected by immunohistochernistry.6.The VEGF-A and HIF-la expressions were determined by Western blotting.7.The proportions of MDSC, TAM (M1/M2), mDC and CD8+T in suspended tumor cells of tumor tissue were detected by FCM.Results1. Compared with A group, the quality of mice and tumor volume of B group and C group grow slowly, the quality of tumor of B group smaller than A group (P<0.05), the measure of tumor suppression rate is 25%; the quality of tumor of C group smaller than A group (P<0.01), the measure of tumor suppression rate is 52%.2.The Microenvironment Density in B group was slightly lower than that in the A group (P<0.05); the Microenvironment Density in C group was deeply lowest in comparison with A group (P<0.01).3.The expressions of VEGF-A in the tumors were decreased, concurrently HIF-la was increased administered with B group compared with A group(P<0.05, respectively). C group alleviated VEGF-A, concurrently elevated HIF-la remarkably better than A group (P<0.01, respectively).4.Flow cytometric data are shown that the proportions of MDSC and TAM (M2 type) were significantly lower, those of mDC and TAM (M1 type) were up-regulated, and CD8+T cells were recruited to infiltrated the tumors by B group, compared with A group (P<0.05, respectively). The changes in Mut-1 antigen loaded DC-T cells combined with Endostar were strongly more than PBS control (P<0.01, respectively).ConclusionsMut-1 antigen loaded DC-T cells combined with Endostar potently strongly inhibited tumor angiogenesis, intensified the hypoxia in the tumors, reduced tumor growth, and efficiently reversed the immunosuppression of the tumor micronment, further exhibited synergistic and much better antitumor effects than monotherapy strategy.