@article{article_1684512, title={Effect of doxorubicin on Caspase-1/GSDMD-mediated pyroptosis in MCF-7 breast cancer spheroids}, journal={Ege Tıp Dergisi}, volume={64}, pages={598–605}, year={2025}, DOI={10.19161/etd.1684512}, url={https://izlik.org/JA66ES67PE}, author={Aydemir, Işıl and Çetindağ, Emre}, keywords={Breast cancer, doxorubicin, spheroid morphology, cytotoxicity}, abstract={Aim: Doxorubicin, a widely used anthracycline antibiotic, exerts its antitumor effects through multiple mechanisms, including DNA intercalation, inhibition of topoisomerase II, and the induction of oxidative stress. This study aimed to evaluate its potential to activate the inflammatory cell death pathway, pyroptosis via Caspase-1 and Gasdermin D (GSDMD), as well as its impact on spheroid morphology in a three-dimensional (3D) cell culture model. Materials and Methods: Spheroids were formed using 3D cell culture techniques and treated with increasing concentrations of doxorubicin (0.1–100 µM) for 24 and 48 hours. Spheroid number and diameter were analyzed using ImageJ v1.47. Caspase-1 and GSDMD protein levels were quantified by ELISA to assess the activation of pyroptotic pathways. Results: Doxorubicin treatment led to a dose-dependent reduction in both spheroid number and size. At 100 µM, spheroid formation was completely inhibited and replaced by disorganized cell clusters. Caspase-1 and GSDMD protein levels increased significantly with higher doxorubicin concentrations, with peak expression observed at 25 µM; suggesting activation of inflammatory cell death pathways. No significant differences were found between 24- and 48-hour treatments. Conclusion: Doxorubicin disrupts 3D spheroid structure and induces the expression of pyroptosis-related proteins in a concentration-dependent manner. These findings highlight its dual role in both impairing tumor architecture and activating inflammatory cell death mechanisms, providing insights into its therapeutic potential in solid tumors.}, number={4}