This study was designed to develop cationic vesicles for doxorubicin (DOX) delivery and to compare anticancer efficacy of these systems uncoated and coated with hyaluronic acid. Cationic nanoformulation was first optimized using various amounts of Span80, DODAB, and cholesterol. The optimized niosomal formulation (CTN4) in terms of vesicle size, surface zeta potential, and colloidal stability was coated with hyaluronic acid and the in vitro therapeutic effectiveness in uterine cervix cancer cells of vesicles loaded with DOX was tested. In vitro studies revealed significantly superior cytotoxicity against Hela cells of niosomes coated with HA compared to uncoated formulations. Moreover, cytotoxicity was also evaluated on normal fibroblast murine cell line, NIH-3T3 cells, and the results obtained demonstrated that HA-coated vesicles exhibited lower cytotoxicity to NIH-3T3 cells compared to uncoated nanovesicles. These findings highlighted how the surface coating influences the effectiveness of niosomes developed as a target drug delivery system and the selectivity and the antitumour efficacy of chemotherapeutic drugs.