1α,25(OH)₂D₃ Radiosensitizes Cancer Cells by Activating the NADPH/ROS Pathway

The radioresistance of tumors affect the outcome of radiotherapy. Accumulating data suggest that 1α,25(OH)₂D₃ is a potential anti-oncogenic molecule in various cancers. In the present study, we investigated the radiosensitive effects and underlying mechanisms of 1α,25(OH)₂D₃ in vitro and in vivo. We found that 1α,25(OH)₂D₃ enhanced the radiosensitivity of lung cancer and ovarian cancer cells by promoting the NADPH oxidase-ROS-apoptosis axis. Compared to the group that only received radiation, the survival fraction and self-renewal capacity of cancer cells treated with a combination of 1α,25(OH)₂D₃ and radiation were decreased. Both apoptosis and ROS were significantly increased in the combination group compared with the radiation only group. Moreover, N-acetyl-L-cysteine, a scavenger of intracellular ROS, reversed the apoptosis and ROS induced by 1α,25(OH)₂D₃, indicating that 1α,25(OH)₂D₃ enhanced the radiosensitivity of cancer cells in vitro by promoting ROS-induced apoptosis. Moreover, our results demonstrated that 1α,25(OH)₂D₃ promoted the ROS level via activating NADPH oxidase complexes, NOX4, p22^phox, and p47^phox. In addition, knockdown of the vitamin D receptor (VDR) abolished the radiosensitization of 1α,25(OH)₂D₃, which confirmed that 1α,25(OH)₂D₃ radiosensitized tumor cells that depend on VDR. Similarly, our study also evidenced that vitamin D₃ enhanced the radiosensitivity of cancer cells in vivo and extended the overall survival of mice with tumors. In summary, these results demonstrate that 1α,25(OH)₂D₃ enhances the radiosensitivity depending on VDR and activates the NADPH oxidase-ROS-apoptosis axis. Our findings suggest that 1α,25(OH)₂D₃ in combination with radiation enhances lung and ovarian cell radiosensitivity, potentially providing a novel combination therapeutic strategy.