Osthole inhibits ovarian carcinoma cells through LC3-mediated autophagy and GSDME-dependent pyroptosis except for apoptosis

Abstract

Ovarian carcinoma (OC) begins in the ovaries and remains a highly lethal malignancy. Despite great efforts have been made to fight against OC, there still remain limited therapeutic options owing to chemotherapy drug resistance and serious side effects. Osthole is a derivative of coumarin and extracted from Cnidium monnieri (L.) Cusson, which has been drawn more attention due to its high biological activity in various disease. However, the underlying mechanism of osthole in OC is still unclear. In this study, we aim to evaluate the mechanism of osthole against OC cells. Methodologically, Cell Counting Kit-8 (CCK-8) and LIVE/DEAD™ Cell Imaging experiments were employed to assess cell viability. 2′,7′-Dichlorofluorescin diacetate (DCFH-DA) staining, flow cytometry, Hoechst staining, JC-1 staining assay and western blotting were performed to study apoptosis. Transmission electron microscopy, western blotting and monodansyl cadaverine (MDC) staining assay were used to study autophagy. Western blotting and microscopy image were employed to determine pyroptosis. Our results demonstrated that osthole could significantly suppress OC cells growth in a dose-dependent manner. We further proved that osthole could inhibit OC cells growth by mitochondria-mediated apoptosis. Meanwhile, we also discovered that osthole could trigger cell autophagy and lead to cell death. Furthermore, our study revealed that osthole could lead to pyroptosis through inducing the cleavage of gasdermin E (c-GSDME) level. Taken together, Osthole could significantly suppress the growth of OC cells and induce OC cells death via apoptosis, pyroptosis and autophagy, which is a promising new drug for the treatment of OC.