Blood vessels are essential for the maintenance and growth of tumor tissues. Furthermore, tumor angiogenesis promotes metastasis, which greatly affects prognosis. Therefore, tumor blood vessels are considered an important target in cancer therapy. Cancer immunotherapy has been developed recently as a new cancer therapeutic. Notably, vaccine therapy with dendritic cells (DCs), which possess potent antigen-presenting capacities, is expected to induce tumor-associated, antigen-specific immunity. In this regard, as tumor endothelial cells (TECs) constituting tumor blood vessels are derived from endothelial cells (ECs) in the host, DC vaccine therapy targeting tumor blood vessels may have applicability in several cancer types. Thus, we attempted to develop DC vaccine therapy that targeted TEC. In our previous studies, in vitro TEC models were created by culturing normal ECs in the culture supernatants of tumor cells. Moreover, we demonstrated that the molecules’ permeability is enhanced in an in vitro TEC model compared with normal ECs. In this study, we examined whether immunotherapy using TEC-extracted proteins as vaccine antigens would be an effective cancer therapy. The results showed that DC vaccine therapy targeting TECs induced anti-tumor effects in a murine Colon-26 solid tumor model and in a lung metastases model using murine B16 melanoma cells. Moreover, anti-angiogenic effects of immunization with TECs were demonstrated. Thus, immunotherapy using the in vitro TEC model as an antigen may be an effective cancer therapeutic. In the future, identifying specific TEC antigens will help generate promising new strategies to inhibit angiogenesis.