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Purpose: Malignant glioma cells are generally insensitive to TRAIL (tumor necrosis factor-related apoptosis-inducing ligand)-induced apoptosis. In this research, we designed a system containing mitoxantrone (MTX) which is loaded into poly lactic-co-glycolic acid (PLGA) nanoparticles (NPs) in order to increase sensitivity of glioblastoma cancer cell to TRAIL plasmid. Methods: PLGA NPs were prepared using a water-in-oil-in-water (W/O/W) double emulsification and solvent evaporation technique and characterized. Synergistic cytotoxicity effect was assessed by both MTT and annexin V-FITC and PI analysis against GL-261 cancer cell line. Result: The combination treatment with PLGA-MTX and TRAIL plasmid showed more cytotoxic effect on GL-261 cancer cell line (cell viability = 16%) compared to MTX-PLGA alone (cell viability 38%) and TRAIL alone (cell viability = 87%). Conclusion: It concluded that PLGA-MTX can be considered a potential agent to sensitize glioblastoma cancer cell to TRAIL. [ABSTRACT FROM AUTHOR]