Chemo‑photothermal therapy for cancer treatment has received increasing attention due to its selective therapeutic effects. In the present study, the anticancer effects of drug‑loaded Fe3O4 magnetic nanoparticles (MNPs) by chemo‑photothermal therapy on U‑87 MG human glioblastoma cells was investigated. Anticancer drug‑loaded Fe3O4 MNPs were prepared by loading temozolomide (TMZ) and indocyanine green (ICG), and were characterized by X‑ray diffraction, UV‑vis spectroscopy, thermal gravimetric analysis, transmission electron microscope, as well as drug‑loading capacity. Following treatment with near‑infrared (NIR) light irradiation, the administration of Fe3O4‑TMZ‑ICG MNPs resulted in the apoptosis of U‑87 MG glioblastoma cells through the generation of reactive oxygen species. Western blot analysis and reverse transcription‑quantitative polymerase chain reaction revealed that Fe3O4‑TMZ‑ICG MNPs with NIR laser irradiation lead to significantly enhanced anticancer effects on U‑87 MG glioblastoma cells through the modulation of intrinsic and extrinsic apoptosis genes, including Bcl‑2‑associated X protein, Bcl‑2, cytochrome c, caspase‑3, Fas associated via death domain and caspase‑8. These results suggest that Fe3O4‑TMZ‑ICG MNPs may be potential candidates when administered as chemo‑phototherapy for the treatment of brain cancer.