Miltirone induces cell death in hepatocellular carcinoma cell through GSDME-dependent pyroptosis

Abstract

Pyroptosis is a form of programmed cell death, and recently described as a new molecular mechanism of chemotherapy drugs in the treatment of tumors. Miltirone, a derivative of phenanthrene-quinone isolated from the root of Salvia miltiorrhiza Bunge, has been shown to possess anti-cancer activities. Here, we found that miltirone inhibited the cell viability of either HepG2 or Hepa1-6 cells, and induced the proteolytic cleavage of gasdermin E (GSDME) in each hepatocellular carcinoma (HCC) cell line, with concomitant cleavage of caspase 3. Knocking out GSDME switched miltirone-induced cell death from pyroptosis to apoptosis. Additionally, the induction effects of miltirone on GSDME-dependent pyroptosis were attenuated by siRNA-mediated caspase three silencing and the specific caspase three inhibitor Z-DEVD-FMK, respectively. Miltirone effectively elicited intracellular accumulation of reactive oxygen species (ROS), and suppressed phosphorylation of mitogen-activated and extracellular signal-regulated kinase (MEK) and extracellular regulated protein kinases 1/2 (ERK1/2) for pyroptosis induction. Moreover, miltirone significantly inhibited tumor growth and induced pyroptosis in the Hepa1-6 mouse HCC syngeneic model. These results provide a new insight that miltirone is a potential therapeutic agent for the treatment of HCC via GSDME-dependent pyroptosis.

A Novel Mechanism of Cannabidiol in Suppressing Hepatocellular Carcinoma by Inducing GSDME Dependent Pyroptosis

“…Consistently, flow cytometry data showed that depletion of GSDME do not rescue the cell death caused by CBD, but GSDME knockdown cells increase the proportion of Annexin V-PE single-positive cells and reduce the percentage of double-positive cells ( Supplementary Figure 6B ). These data indicated that the deficiency of GSDME may switch CBD-triggered cell death from pyroptosis to apoptosis ( Zhang et al, 2020 ). Previous studies have demonstrated that GSDME-dependent pyroptosis can be activated by pro-apoptotic caspases, such as caspase-3 25 .…”

“…Meanwhile, recent research has shown that chemotherapy-induced pyroptosis is mediated by the BAK/BAX–caspase-3–GSDME pathway ( Zhang et al, 2018 ), indicating that the pro-apoptotic factors of the BCL-2 family may be involved in cell pyroptosis. Moreover, Zhang et al (2020) have demonstrated that mitochondrial dysfunction triggered caspase3–GSDME pathway activation can eventually lead to pyroptosis in response to Miltirone in HCC cells. There may therefore be a similar mechanism in CBD-induced pyroptosis.…”

Caspase 3/GSDME-dependent pyroptosis contributes to chemotherapy drug-induced nephrotoxicity

“…More recently, many studies have indicated that reagents or drugs targeting GSDME showed an antitumor effect. For instance, Miltirone, derived from a traditional herb Salvia miltiorrhiza, was shown to possess antitumor activity by inducing GSDME activation in hepatocellular carcinoma 51 . Furthermore, a PLK1 kinase inhibitor was reported to improve the effect of cisplatin in the treatment of esophageal squamous cell carcinoma by inducing pyroptosis 52 .…”

Prognostic and Immunological Role of Gasdermin E in Pan-Cancer Analysis

“…Gasdermin proteins have been documented to possess inherent toxic activity of the Gasdermin-N terminal fragments, which are frequently masked by inhibitory Gasdermin-C terminal fragments (19,44,45). Proteolytic cleavage the hinge loop between these terminals to unleash the necrotic Gasdermin-N fragments to squeeze the inner leaflet of membrane and form nanopores with a size of~18 nm in the cellular plasma membrane (44,(46)(47)(48). Physical rupture of pyroptotic cells causes the leakage of inflammatory factors IL-18, IL-1b, and DAMPs to amplify the local and systemic inflammatory response, suggesting the immunogenic potential of pyroptosis (49,50).…”