The present invention discloses a short peptide targeting EPS8 binding with EGFR and use thereof, and sequence of the short peptide is N′-Arg-Lys-Lys-Asn-Lys-Pro-Pro-Pro-Pro-Lys-Lys-C′. The short peptide can effectively inhibit proliferation of EPS8 positive tumors, and can also be used to make a pharmaceutical preparation for treating EPS8 positive tumors, which has the potential of being developed into anti-cancer peptide inhibitor drugs.