Biomarker-specific photothermal nanoparticles that can efficiently sense the markers, which are overexpressed in distinguished adenocarcinomas, has attracted much interest in an aspect of efficacy increase of cancer treatment. In this study, we demonstrated a promising prospect of smart photothermal therapy agent employing anti-epidermal growth factor receptor aptamer (AptEGFR)-conjugated polyethylene glycol (PEG)layted gold nanorods (AptEGFR-PGNRs). The cetyltrimethylammonium bromide bilayer on GNRs was replaced with heterobifunctional polyethylene glycol (COOHPEG- SH) not only to serve as a biocompatible stabilizer and but also to conjugate AptEGFR. Subsequently, to direct photothermal therapy agent toward epithelial cancer cells, the carboxylated PEGylated GNRs (PGNRs) were further functionalized with AptEGFR using carbodiimide chemistry. And then, to assess the potential as biomarker-specific photothermal therapy agent of synthesized AptEGFR-PGNRs, the optical properties, biocompatibility, colloidal stability, binding affinity and epicellial cancer cell killing efficacy in vitro/in vivo under NIR laser irradiation were investigated. As a results, AptEGFR-PGNRs exhibit excellent tumor targeting ability and feasibility of effective photothermal ablation cancer therapy.