Polyethylene glycol-poly(lactic-co-glycolic acid) (PEG-PLGA) nanoparticles coated with neutrophil membranes were fabricated to afford a bio-mimicking delivery system and achieve targeted delivery of chemotherapeutics towards malignant melanoma via systemic administration. Using celastrol as the model compound, celastrol-loaded PEG-PLGA nanoparticles coated with neutrophil membranes displayed significantly enhanced cytotoxicity and apoptosis rate in a murine melanoma cell line B16F10 compared to celastrol-loaded PEG-PLGA nanoparticles. Moreover, PEG-PLGA nanoparticles coated with neutrophil membranes exhibited significantly higher internalization efficiency in B16F10 cells than nanoparticles without membrane coating. Next, a B16F10 tumor xenograft mice model was established to explore the biodistribution profiles of PEG-PLGA nanoparticles coated with neutrophil membranes which showed remarkably prolonged blood circulation and more selective accumulation at the tumor site. Celastrol-loaded PEG-PLGA nanoparticles coated with neutrophil membranes also demonstrated greatly improved antitumor efficacy in B16F10 tumor bearing mice xenografts. Taken together, PEG-PLGA nanoparticles coated with neutrophil membranes represent a highly promising nanoscale delivery system to achieve tumor-targeted therapy following systemic administration.