OBJECTIVE: To prepare peptide T7-modified vincristine sulfate (VCR)-apoferritin (APO) nanoparticles, and to study its in vitro targeting to tumor cells. METHODS: VCR-APO nanoparticles were prepared by dissociation-recombination method. The targeting peptide T7 was modified to prepare T7-VCR-APO nanoparticles. Target attachment efficiency was evaluated by MS analysis and UV spectrophotometry. The morphology, particle size, Zeta potential and encapsulation efficiency of prepared nanoparticles were all characterized. Using glioma cells C6 as model, the targeting effects of T7-VCR-APO nanoparticles on C6 cells and tumor spheres were investigated. The uptake of T7-VCR-APO nanoparticles by C6 cells was investigated under low temperature and by 4 kinds of cell endocytosis inhibitors. RESULTS: T7 peptide was confirmed to be linked to nanoparticles by MS, with attachment rate of 68.39%. The prepared T7-VCR-APO nanoparticles were round and uniform. The particle size was (31.14±1.26) nm, Zeta potential was (-23.30±0.42) mV, and entrapment efficiency was (39.49±2.84)%. The in vitro targeting study of tumor cells showed that T7-VCR-APO nanoparticles could be effectively taken up by C6 cells and spheres, and can reach the sphere center. The uptake of nanoparticles by C6 cells was the result of a multi-pathway interaction mainly through caveolin-mediated endocytosis and concomitant macrophage pathways. CONCLUSIONS: T7-VCR-APO nanoparticles are prepared successfully, can achieve targeting drug delivery to glioma C6 cells in vitro,and endocytosis efficiency is greatly improved.