DZ-1-artesunate conjugate induces mitochondria-mediated, reactive oxygen species-dependent apoptosis in colorectal cancer tumoroids.

We previously reported that the heptamethine cyanine dye-conjugated artesunate (DZ-1-ART) induces apoptosis in monolayer (2D) cell culture models. However, in 2D cultures, cells grow on flat, rigid plastic surfaces that fail to recapitulate the three-dimensional architecture of tumor tissues. This artificial environment alters cell polarity, morphology, and mechanical signaling, leading to non-physiological behavior. To overcome these limitations, we developed patient-derived tumoroids to assess the tumoricidal efficacy of the DZ-1-ART conjugate. In this study, tumoroids were established from fresh tissue samples of a malignant neoplasm of the sigmoid colon. The anticancer activity of DZ-1-ART was evaluated in these tumoroids. Propidium iodide staining confirmed DZ-1-ART-induced cytotoxicity, while TUNEL and immunoblotting assays demonstrated that this cytotoxicity was mediated by apoptosis. Furthermore, MitoTracker staining and near-infrared fluorescence indicated mitochondrial localization of DZ-1-ART. The JC-1 assay showed disruption of mitochondrial membrane potential following DZ-1-ART treatment. Additionally, deferoxamine and MitoTEMPO pretreatment revealed that DZ-1-ART induced mitochondria-mediated reactive oxygen species (ROS) generation in tumoroids. Collectively, these findings suggest that DZ-1-ART acts as a potent mitochondria-targeting anticancer agent with potential for precision therapy.