Ribosome-inactivating proteins (RIPs) from plants have emerged as potent bioactive molecules with diverse therapeutic potential, including anticancer activity. In this study, we isolated and characterized RIPs from Cucurbita moschata (pumpkin). The proteins were purified using a combination of precipitation and chromatographic methods and characterized by SDS-PAGE and enzymatic assays. We assessed their cytotoxic effects on plant bioassays, revealing significant anticancer potential. These findings suggest that Cucurbita moschata RIPs could be valuable candidates for targeted anticancer therapies.