The invention belongs to the technical field of polypeptide in the biological chemistry filed and particularly relates to an FAP source anti-tumor CTL epitope peptide P265 and application thereof in preparation of tumor therapeutic peptide vaccine. The anti-tumor CTL epitope peptide P265 is nonapeptide, the molecular weight is 1,040.18, the specific sequence is FIIDTTYPA. More than 90% of specific antigen FAP which has high expression in epithelial ovarian tumor and almost no expression in normal tissue is used, HLA-A2 restrictive CTL epitope of the FAP is predicated and screened according to the primary structure of the FAP, it is determined that the CTL epitope peptide and the HLA-A2 molecules can produce high combining capacity, specificity CTL reaction can be effectively activated, and the FAP source anti-tumor CTL epitope peptide P265 has the feasibility of becoming the therapeutic tumor peptide vaccine.