In this study, an HER2 aptamer-decorated curcumin-loaded human serum albumin nanoparticle (Apt-HSA/CCM NP) was developed and characterized as a new anticancer formulation for targeted delivery to human epithelial growth factor receptor 2 (HER2) overexpressing breast cancer cells. Conjugation of HER2 Apt to the surface of HSA NPs was confirmed by gel electrophoresis and FTIR analysis. The obtained NPs have the hydrodynamic diameter of 281.1 ± 11.1 nm and zeta potential of -33.3 ± 2.5 mV. The data demonstrated that encapsulation of curcumin in HSA NPs by desolvation method has increased water solubility by 400 folds. Fluorescent microscopy image demonstrated remarkable cytoplasmic uptake of Apt-HSA/CCM NPs in HER2-overexpressing SK-BR-3 cells compared to unconjugated counterparts. Cytotoxicity experiments demonstrated no significant difference between cytotoxic effect of free curcumin and non-targeted HSA/CCM NPs in both HER2 positive and HER2 negative cell lines. However, the toxicity of Apt-HSA/CCM NPs was significantly higher and cell viability reached 36% after 72 h in SK-BR3 cell line. These results suggest that this targeted delivery system has the potential to be considered as a promising candidate for the treatment of HER2 positive cancer cells.