The invention discloses a fusion protein capable of inducing and activating cancer targeting T-cells as well as a preparation method and application of the fusion protein. The fusion protein contains peptide acting with cancer cells as well as costimulatory molecules B7.1, wherein the peptide acting with the cancer cells is selected from a transforming growth factor-alpha, an epidermal growth factor, a vascular endothelial growth factor, gonadotropinreleasing hormone or gastrin-releasing peptide. The fusion protein has a targeting function; the fusion protein can act with VEGFR, EGFR, GnRH-R, or GRP-R respectively on one hand and interact with corresponding receptors CD28 and CTLA-4 expressed on the T-cells, therefore the targeting of the T-cells is positioned at the periphery of the cancer cells greatly expressing VEGFR, EGFR, GnRH-R, or GRP-R. Proven by experiments, the fusion protein can restrain the growth of tumors and cause cancer cells to apoptosis.