The invention belongs to the field of technical methods for preventing tumors, and in particular relates to a tumor microenvironment induced cancerous human umbilical vein endothelial cell vaccine andan application thereof in resisting angiogenesis of colon carcinoma. The vaccine is prepared by the following steps: collecting supernatant of CT26 cells, inducing HUVEC cells via the CT26 cell supernatant, collecting the HUVEC cells, immobilizing glutaraldehyde and the like. According to the invention, tumor vessel endothelial cells function as a target in tumor immnuotherapy, belonging to active immunotherapy. The cancerous HUVEC vaccine is prepared through inducing by an in-vitro simulated tumor microenvironment. Based upon preliminary experimental results, it is indicated that the prepared cancerous HUVEC vaccine can cause a more effective action in inhibiting tumor angiogenesis, and subsequently, an effect of resisting colon carcinoma growth can be developed; therefore, a relativelygood application effect can be achieved; and meanwhile, a new possibility is also provided for the application of a treatment method based on resisting tumor angiogenesis.