Phosphorylated IRF3 promotes GSDME-mediated pyroptosis through RIPK1/FADD/caspase-8 complex formation during mitotic arrest in ovarian cancer

Abstract Inducing mitotic arrest with anti-mitotic drugs is an effective strategy for cancer therapy. However, the ultimate fate of cells that undergo prolonged mitotic arrest remains largely uncertain. In this study, paclitaxel and nocodazole were used to induce prolonged mitotic arrest in ovarian...

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Main Authors: Wenjian Gong, Dongchen Zhou, Qiuyang Xu, Linghui Wang, Mengshi Luo, Yuewen Zhang, Zhiqi Liao, Fan Xiong, Guangnian Zhao, Bingbing Zhao, Qinglei Gao, Yong Fang
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Cell Communication and Signaling
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Online Access:https://doi.org/10.1186/s12964-025-02322-9
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author Wenjian Gong
Dongchen Zhou
Qiuyang Xu
Linghui Wang
Mengshi Luo
Yuewen Zhang
Zhiqi Liao
Fan Xiong
Guangnian Zhao
Bingbing Zhao
Qinglei Gao
Yong Fang
author_facet Wenjian Gong
Dongchen Zhou
Qiuyang Xu
Linghui Wang
Mengshi Luo
Yuewen Zhang
Zhiqi Liao
Fan Xiong
Guangnian Zhao
Bingbing Zhao
Qinglei Gao
Yong Fang
author_sort Wenjian Gong
collection DOAJ
description Abstract Inducing mitotic arrest with anti-mitotic drugs is an effective strategy for cancer therapy. However, the ultimate fate of cells that undergo prolonged mitotic arrest remains largely uncertain. In this study, paclitaxel and nocodazole were used to induce prolonged mitotic arrest in ovarian cancer cells, triggering mitotic catastrophe, during which these cells exhibited hallmarks of pyroptosis. Subsequently, small interfering RNA (siRNA)-mediated downregulation of Gasdermin E (GSDME) inhibited pyroptosis, suggesting that GSDME plays an essential role in this process. The upstream signaling pathway was further investigated through caspase-3 inhibition and caspase-8 knockdown, which demonstrated that pyroptosis induced by paclitaxel and nocodazole was mediated by the caspase-8/caspase-3/GSDME pathway. Moreover, during mitotic arrest, phosphorylation of IRF3, mediated by cGAS/TBK1, led to the formation of the RIPK1/FADD/caspase-8 complex, which subsequently activated caspase-8 and initiated downstream GSDME-mediated pyroptosis. Knockdown of components of this complex or mutation of the IRF3 phosphorylation site inhibited pyroptosis. Furthermore, in vivo experiments also demonstrated that paclitaxel inhibited tumor growth by inducing GSDME-mediated pyroptosis and activating the anti-tumor immune infiltration. TCGA data further suggested that ovarian cancer cases treated with paclitaxel, showing high expression of GSDME and caspase-3, exhibited a more favorable tumor immune microenvironment. This study not only elucidated the specific mechanism of pyroptosis mediated by phosphorylated IRF3 during prolonged mitotic arrest but also revealed that mitotic arrest-induced pyroptosis could enhance immune infiltration in ovarian cancer, providing valuable insights for clinical treatment strategies.
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spelling doaj-art-68bf155dd492443e8e94dc585a96bbba2025-08-20T04:01:35ZengBMCCell Communication and Signaling1478-811X2025-07-0123111710.1186/s12964-025-02322-9Phosphorylated IRF3 promotes GSDME-mediated pyroptosis through RIPK1/FADD/caspase-8 complex formation during mitotic arrest in ovarian cancerWenjian Gong0Dongchen Zhou1Qiuyang Xu2Linghui Wang3Mengshi Luo4Yuewen Zhang5Zhiqi Liao6Fan Xiong7Guangnian Zhao8Bingbing Zhao9Qinglei Gao10Yong Fang11Department of Gynecological Oncology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Gynecological Oncology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Gynecological Oncology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Gynecological Oncology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Gynecological Oncology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Gynecological Oncology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Gynecological Oncology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Gynecological Oncology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Gynecological Oncology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Obstetrics and Gynecology, Guangxi Medical University of Cancer HospitalDepartment of Gynecological Oncology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and TechnologyDepartment of Gynecological Oncology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and TechnologyAbstract Inducing mitotic arrest with anti-mitotic drugs is an effective strategy for cancer therapy. However, the ultimate fate of cells that undergo prolonged mitotic arrest remains largely uncertain. In this study, paclitaxel and nocodazole were used to induce prolonged mitotic arrest in ovarian cancer cells, triggering mitotic catastrophe, during which these cells exhibited hallmarks of pyroptosis. Subsequently, small interfering RNA (siRNA)-mediated downregulation of Gasdermin E (GSDME) inhibited pyroptosis, suggesting that GSDME plays an essential role in this process. The upstream signaling pathway was further investigated through caspase-3 inhibition and caspase-8 knockdown, which demonstrated that pyroptosis induced by paclitaxel and nocodazole was mediated by the caspase-8/caspase-3/GSDME pathway. Moreover, during mitotic arrest, phosphorylation of IRF3, mediated by cGAS/TBK1, led to the formation of the RIPK1/FADD/caspase-8 complex, which subsequently activated caspase-8 and initiated downstream GSDME-mediated pyroptosis. Knockdown of components of this complex or mutation of the IRF3 phosphorylation site inhibited pyroptosis. Furthermore, in vivo experiments also demonstrated that paclitaxel inhibited tumor growth by inducing GSDME-mediated pyroptosis and activating the anti-tumor immune infiltration. TCGA data further suggested that ovarian cancer cases treated with paclitaxel, showing high expression of GSDME and caspase-3, exhibited a more favorable tumor immune microenvironment. This study not only elucidated the specific mechanism of pyroptosis mediated by phosphorylated IRF3 during prolonged mitotic arrest but also revealed that mitotic arrest-induced pyroptosis could enhance immune infiltration in ovarian cancer, providing valuable insights for clinical treatment strategies.https://doi.org/10.1186/s12964-025-02322-9Ovarian cancerPyroptosisMitotic arrestIRF3
spellingShingle Wenjian Gong
Dongchen Zhou
Qiuyang Xu
Linghui Wang
Mengshi Luo
Yuewen Zhang
Zhiqi Liao
Fan Xiong
Guangnian Zhao
Bingbing Zhao
Qinglei Gao
Yong Fang
Phosphorylated IRF3 promotes GSDME-mediated pyroptosis through RIPK1/FADD/caspase-8 complex formation during mitotic arrest in ovarian cancer
Cell Communication and Signaling
Ovarian cancer
Pyroptosis
Mitotic arrest
IRF3
title Phosphorylated IRF3 promotes GSDME-mediated pyroptosis through RIPK1/FADD/caspase-8 complex formation during mitotic arrest in ovarian cancer
title_full Phosphorylated IRF3 promotes GSDME-mediated pyroptosis through RIPK1/FADD/caspase-8 complex formation during mitotic arrest in ovarian cancer
title_fullStr Phosphorylated IRF3 promotes GSDME-mediated pyroptosis through RIPK1/FADD/caspase-8 complex formation during mitotic arrest in ovarian cancer
title_full_unstemmed Phosphorylated IRF3 promotes GSDME-mediated pyroptosis through RIPK1/FADD/caspase-8 complex formation during mitotic arrest in ovarian cancer
title_short Phosphorylated IRF3 promotes GSDME-mediated pyroptosis through RIPK1/FADD/caspase-8 complex formation during mitotic arrest in ovarian cancer
title_sort phosphorylated irf3 promotes gsdme mediated pyroptosis through ripk1 fadd caspase 8 complex formation during mitotic arrest in ovarian cancer
topic Ovarian cancer
Pyroptosis
Mitotic arrest
IRF3
url https://doi.org/10.1186/s12964-025-02322-9
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