Osthole Induces Apoptosis and Caspase-3/GSDME-Dependent Pyroptosis via NQO1-Mediated ROS Generation in HeLa Cells

Abstract

Osthole is a natural coumarin which has been proved to inhibit growth of cancer cells by inducing cell death, while its mechanism was considered to be just caused by apoptosis. In our study, we found that osthole activated not just apoptosis, but also pyroptosis which is a form of regulated cell death accompanied by loss of cell membrane integrity and lactate dehydrogenase (LDH) release. Caspase-3 is a key protein of apoptosis as well as pyroptosis. The apoptosis and pyroptosis induced by osthole were all inhibited by irreversible caspase-3 inhibitor Z-DEVD-FMK. Meanwhile, knockdown of gasdermin E (GSDME) only reduced the osthole-induced pyroptosis but did not affect the occurrence of apoptosis. Our proteomic analysis revealed that the expression of NAD(P)H: quinone oxidoreductase 1 (NQO1) was decreased in osthole-treated cells. Moreover, NQO1 inhibition by osthole induced the overproduction of reactive oxygen species (ROS), as well as apoptosis and pyroptosis. ROS inhibitor N-Acetyl-L-cysteine (NAC) not only reduced osthole-induced apoptosis but also reversed its effect on the pyroptosis. Meanwhile, knockdown of NQO1 by si-NQO1 or its inhibitor dicoumarol (DIC) not only enhanced ROS generation but also strengthened the GSDME-mediated pyroptosis. Finally, we demonstrated that osthole inhibited tumor growth and the expression of NQO1 in a HeLa xenograft mode. Similar to the results in vitro, osthole stimulated the activation of caspase-3, PARP, and GSDME in vivo. Taken together, all these data suggested that osthole induced apoptosis and caspase-3/GSDME-mediated pyroptosis via NQO1-mediated ROS accumulation.

Caspase-3/GSDME dependent pyroptosis contributes to offspring lung injury induced by gestational PFOS exposure via PERK/ATF4 signaling

“…Prior research has established that the level of GSDME plays a crucial role in determining which form of cell death could be induced (Wang et al 2017 ). Cells with elevated GSDME expression undergo pyroptosis upon caspase-3 activation, while cells with low GSDME expression undergo apoptosis (Liu et al 2022 ; Wang et al 2022a ; Yu et al 2019 ). In line with these findings, our study demonstrated that PFOS primarily induced pyroptosis by up-regulation of GSDME.…”

Pyroptosis and its role in gynecologic cancer

“…It is capable of inhibiting cell proliferation, causing DNA damage, inducing ROS generation, and leading to pyroptosis [22]. Extracted from plants, osthole is also a natural compound with anticancer effects [23]. The reactive oxygen species (ROS) in the osthole-treated cells accumulate due to the decrease of NAD(P)H quinone oxidoreductase 1 (NQO1).…”

Pyroptosis in glioma: Current management and future application

“…77 Osthole, a naturally occurring coumarin, has also been found to induce pyroptosis mediated by caspase-3/GSDME. 78 Alantolactone (ATL), a terpenoid derived from the traditional Chinese medicinal herb Inula helenium L., exhibits notable anti-inflammatory, antibacterial, and antitumor properties. ATL has been found to induce simultaneous apoptosis and GSDME-dependent pyroptosis through the activation of caspase 9 and caspase 3, which is mediated by ROS and mitochondria.…”

“…Moreover, NBT activates caspase‐3 in esophageal cancer cells, causing GSDME‐mediated pyroptosis 77 . Osthole, a naturally occurring coumarin, has also been found to induce pyroptosis mediated by caspase‐3/GSDME 78 . Alantolactone (ATL), a terpenoid derived from the traditional Chinese medicinal herb Inula helenium L., exhibits notable anti‐inflammatory, antibacterial, and antitumor properties.…”