Mitoxantrone (MIT) is a chemotherapeutic agent with promising anticancer efficacy. In this study, Pluronic F68-vitamine E succinate (F68-VES) amphiphilic polymer micelles were developed for delivering MIT and enhancing its anticancer activity. MIT-loaded F68-VES (F68-VES/MIT) micelles were prepared via the solvent evaporation method with self-assembly under aqueous conditions. F68-VES/MIT micelles were found to be of optimal particle size with the narrow size distribution. Transmission electron microscopy images of F68-VES/MIT micelles showed homogeneous spherical shapes and smooth surfaces. F68-VES micelles had a low critical micelle concentration value of 3.311 mg/L, as well as high encapsulation efficiency and drug loading. Moreover, F68-VES/MIT micelles were stable in the presence of fetal bovine serum for 24 hours and maintained sustained drug release in vitro. Remarkably, the half maximal inhibitory concentration (IC) value of F68-VES/MIT micelles was lower than that of free MIT in both MDA-MB-231 and MCF-7 cells (two human breast cancer cell lines). In addition, compared with free MIT, there was an increased trend of apoptosis and cellular uptake of F68-VES/MIT micelles in MDA-MB-231 cells. Taken together, these results indicated that F68-VES polymer micelles were able to effectively deliver MIT and largely improve its potency in cancer therapy.