The prognosis for glioblastoma multiforme (GBM), the adult brain and spinal cord tumour with the poorest prognosis, has not improved enough with current therapies. Investigating potential molecular routes of tumour growth and metastasis and novel therapeutic methods are, therefore, obviously necessary. Although some natural substances have been shown to increase intracellular Ca²⁺ concentration and ROS generation by activating TRPM2 channel and potentiate the anticancer effects of doxorubicin (DOX), and chrysin (CHR) has been shown to enhance the anticancer effect of DOX, its anticancer effect through TRPM2-mediated Ca²⁺ homeostasis and ROS generation in glioblastoma tumour cells (DBTRG) has not been investigated. This work aimed to clarify the molecular processes behind the synergistic chemotherapeutic action of DOX and CHR in DBTRG cells. For this purpose, we investigated the stimulatory role of CHR on DOX-induced cell death via TRPM2 channel activation in DBTRG cells. DOX treatment caused cell cytotoxicity, increased apoptosis (caspase 3 and 9), ROS, lipid peroxidation levels, and decreased cell viability and antioxidant levels. The combination of CHR and dOx made the treatment even more effective. The effects in cells were reduced with treatments with 2APB, a TRPM2 antagonist. In conclusion, the increase in ROS and cell death levels mediated by TRPM2 activation in DOX-induced DBTRG cells was further enhanced by CHR treatment. The combination of CHR and DOX can be used as a successful agent in the treatment of glioblastoma tumours.