Cordyceps militaris extract induces apoptosis and pyroptosis via caspase‐3/PARP/GSDME pathways in A549 cell line

Abstract

Cordyceps militaris (CM) is traditionally used as dietary therapy for lung cancer patients in China. CM extract (CME) is hydrosoluble fraction of CM and extensively investigated. Caspase‐3‐involved cell death is considered as its major anticancer mechanism but inconclusive. Therefore, we explore its caspase‐3‐dependent programmed cell death nature (apoptosis and pyroptosis) and validate its caspase‐3‐dependent property in loss‐of‐function experiment. Component profile of CME is detected by High Performance Liquid Chromatography‐ quadrupole time‐of‐flight mass spectrometry (HPLC‐qTOF). Results show that CME causes pyroptosis‐featured cell bubbling and cell lysis and inhibits cell proliferation in A549 cell. CME induces chromatin condensing and makes PI+/annexin V+ staining in bubbling cells, indicating genotoxicity, apoptosis, and pyroptosis cell death are caused by CME. High concentration of CME (200 μg/ml) exerts G2/M and G0 cell cycles arresting and suppresses P53‐downstream proliferative proteins, including P53, P21, CDC25B, CyclinB1, Bcl‐2, and BCL2 associated agonist of cell death (BAD), but 1–100 μg/ml of CME show less effect on proteins above. Correspondingly, caspase‐3 activity and caspase‐3 downstream proteins including pyroptotic effector gasdermin‐E (GSDME) and apoptotic marker cleaved‐poly‐ADP‐ribose polymerase (PARP) are significantly promoted by CME. Moreover, regarding membrane pore formation in pyroptotic cell, expression of membrane GSDME (GSDME antibody conjugated with PE‐Cy7 for detection in flow cytometry) is remarkably increased by CME treatment. By contrast, other pyroptosis‐related proteins such as P2X7, NLRP3, GSDMD, and Caspase‐1 are not affected after CME treatment. Additionally, TET2 is unexpectedly raised by CME. In present of caspase‐3 inhibitor Ac‐DEVD‐CHO (Ac‐DC), CME‐induced cytotoxicity, cell bubbling, and genotoxicity are reduced, and CME‐induced upregulation of apoptosis (cleaved‐PARP‐1) and pyroptosis (GSDME‐NT) proteins are reversed. Lastly, 22 components are identified in HPLC‐qTOF experiment, and they are classified into trophism, neoadjuvant component, cytotoxic component, and cancer deterioration promoter according to previous references. Conclusively, CME causes caspase‐3‐dependent apoptosis and pyroptosis in A549 through caspase‐3/PARP and caspase‐3/GSDME pathways, and it provides basic insight into clinic application of CME for cancer patients.

Encapsulation of Selenium Nanoparticles and Metformin in Macrophage-Derived Cell Membranes for the Treatment of Spinal Cord Injury

“…56,57 The caspase-3 apoptosis pathway plays a key role in apoptosis. 58 In the present study, immunofluorescence and Tunel assays showed that SeNPs-Met-MVs could significantly reduce the expression of apoptotic factors (cleaved caspase-3). At the same time, SeNPs-Met-MVs promoted the recovery of motor function in previous SCI.…”

“…Previous studies have shown that oxidative stress ultimately leads to apoptosis of neuronal cells . The cysteine protease family is the primary regulator of apoptosis. , The caspase-3 apoptosis pathway plays a key role in apoptosis . In the present study, immunofluorescence and Tunel assays showed that SeNPs-Met-MVs could significantly reduce the expression of apoptotic factors (cleaved caspase-3).…”

The multifaceted roles of GSDME-mediated pyroptosis in cancer: therapeutic strategies and persisting obstacles

“…In addition, GSDME interacts physically with EGFR, covering the EGFRY1045 site and contributing to its stability [ 107 ]. The hydrosoluble fraction of cordyceps militaris extract (CME), a dietary herb for lung cancer patients, has been shown to induce A549 cell death by simultaneously activating caspase-3/GSDME pyroptotic pathways and caspase-3/PARP apoptotic pathways [ 108 ]. Yu et al revealed that the pyroptosis inhibitor (DSF) and apoptosis inhibitor (Z-VAD-FMK) prevented the cell mortality induced by nitidine chloride [ 109 ].…”

Pyroptosis and Insulin Resistance in Metabolic Organs

“…Also the newly discovered key protein of pyroptosis GSDME primarily relies on caspase-3 to mediate pyroptosis, and this pathway is of relevance especially in cancer. In the field of cancer treatment, chemotherapeutic drugs can mediate pyroptosis through the caspase-3/GSDME pathway, thereby enhancing the effect of chemotherapy; for instance, a Cordyceps militaris extract can induce caspase-3-dependent pyroptosis in A549 cells through caspase-3/PARP and caspase-3/GSDME pathways [ 45 ]. It has been reported that pathogenic Yersinia can lead to caspase-8-dependent pyroptosis after entering cells by inhibiting transforming growth factor β-activated kinase 1, activating and cleaving receptor-interacting protein kinase 1 [ 46 ] ( Figure 1 ).…”