{"title":"嗜中性粒细胞胞外陷阱通过丝氨酸-1表达促进胃癌上皮-间质转化的分子机制","authors":"Zhen Ma, Xiaolin Li, Shifeng Yang, Hao Yang, Ange Zhang, Nana Li, Xiaoming Zou","doi":"10.1002/jbt.70157","DOIUrl":null,"url":null,"abstract":"<p>Gastric cancer remains a significant global health concern, with its progression and metastasis often associated with epithelial–mesenchymal transition (EMT). This study investigated the role of neutrophil extracellular traps (NETs) in promoting gastric cancer EMT by regulating SERPINE-1 expression, which encodes plasminogen activator inhibitor-1 (PAI-1). Western blot and immunohistochemistry were used to detect protein expression. Cell Counting Kit-8 was tested for cell proliferation ability using clones. The SERPINE-1 gene was knocked down using lentivirus. Immunofluorescence was used to detect the co-expression of proteins, and a Transwell assay and wound-healing assay were used to investigate the migration ability of cells. Experimental conclusions were verified in vivo using a nude mouse model. We first demonstrated overexpression of PAI-1 in gastric cancer tissues and cell lines. Subsequently, we found that NETs significantly enhanced the expression of EMT-related markers. These changes were accompanied by increases in cell invasion, migration, proliferation and tumour sphere formation. To further elucidate the mechanism, we employed lentivirus-mediated SERPINE-1 knockdown to reverse NET-induced EMT phenotype effectively. Mechanistically, we found that NETs activated the transforming growth factor (TGF)-β signalling pathway via PAI-1 as evidenced by increased expression of TGF-β1, TGF-βR1, TGF-βR2, phosphorylated Smad2/3 and Smad4. Finally, in vivo experiments using a nude mouse model of gastric cancer liver metastasis confirmed that NET-treated HGC-27 cells exhibited enhanced metastatic potential and SERPINE-1 knockdown abrogated metastatic potential. Our findings reveal a novel mechanism by which NETs promote EMT and metastasis in gastric cancer via the PAI-1–TGF-β axis. PAI-1 can be used as a potential target for the treatment of gastric cancer, and the expression of PAI-1 is closely related to the prognosis of patients with gastric cancer. Therapeutic strategies targeting NETs or PAI-1 may help prevent EMT and metastasis of gastric cancer and improve clinical outcomes in patients.</p>","PeriodicalId":15151,"journal":{"name":"Journal of Biochemical and Molecular Toxicology","volume":"39 3","pages":""},"PeriodicalIF":3.2000,"publicationDate":"2025-03-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jbt.70157","citationCount":"0","resultStr":"{\"title\":\"Molecular Mechanisms of Neutrophil Extracellular Traps in Promoting Gastric Cancer Epithelial–Mesenchymal Transition Through SERPINE-1 Expression\",\"authors\":\"Zhen Ma, Xiaolin Li, Shifeng Yang, Hao Yang, Ange Zhang, Nana Li, Xiaoming Zou\",\"doi\":\"10.1002/jbt.70157\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Gastric cancer remains a significant global health concern, with its progression and metastasis often associated with epithelial–mesenchymal transition (EMT). This study investigated the role of neutrophil extracellular traps (NETs) in promoting gastric cancer EMT by regulating SERPINE-1 expression, which encodes plasminogen activator inhibitor-1 (PAI-1). Western blot and immunohistochemistry were used to detect protein expression. Cell Counting Kit-8 was tested for cell proliferation ability using clones. The SERPINE-1 gene was knocked down using lentivirus. Immunofluorescence was used to detect the co-expression of proteins, and a Transwell assay and wound-healing assay were used to investigate the migration ability of cells. Experimental conclusions were verified in vivo using a nude mouse model. We first demonstrated overexpression of PAI-1 in gastric cancer tissues and cell lines. Subsequently, we found that NETs significantly enhanced the expression of EMT-related markers. These changes were accompanied by increases in cell invasion, migration, proliferation and tumour sphere formation. To further elucidate the mechanism, we employed lentivirus-mediated SERPINE-1 knockdown to reverse NET-induced EMT phenotype effectively. Mechanistically, we found that NETs activated the transforming growth factor (TGF)-β signalling pathway via PAI-1 as evidenced by increased expression of TGF-β1, TGF-βR1, TGF-βR2, phosphorylated Smad2/3 and Smad4. Finally, in vivo experiments using a nude mouse model of gastric cancer liver metastasis confirmed that NET-treated HGC-27 cells exhibited enhanced metastatic potential and SERPINE-1 knockdown abrogated metastatic potential. Our findings reveal a novel mechanism by which NETs promote EMT and metastasis in gastric cancer via the PAI-1–TGF-β axis. PAI-1 can be used as a potential target for the treatment of gastric cancer, and the expression of PAI-1 is closely related to the prognosis of patients with gastric cancer. Therapeutic strategies targeting NETs or PAI-1 may help prevent EMT and metastasis of gastric cancer and improve clinical outcomes in patients.</p>\",\"PeriodicalId\":15151,\"journal\":{\"name\":\"Journal of Biochemical and Molecular Toxicology\",\"volume\":\"39 3\",\"pages\":\"\"},\"PeriodicalIF\":3.2000,\"publicationDate\":\"2025-03-10\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jbt.70157\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Biochemical and Molecular Toxicology\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/jbt.70157\",\"RegionNum\":3,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Biochemical and Molecular Toxicology","FirstCategoryId":"3","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/jbt.70157","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
Molecular Mechanisms of Neutrophil Extracellular Traps in Promoting Gastric Cancer Epithelial–Mesenchymal Transition Through SERPINE-1 Expression
Gastric cancer remains a significant global health concern, with its progression and metastasis often associated with epithelial–mesenchymal transition (EMT). This study investigated the role of neutrophil extracellular traps (NETs) in promoting gastric cancer EMT by regulating SERPINE-1 expression, which encodes plasminogen activator inhibitor-1 (PAI-1). Western blot and immunohistochemistry were used to detect protein expression. Cell Counting Kit-8 was tested for cell proliferation ability using clones. The SERPINE-1 gene was knocked down using lentivirus. Immunofluorescence was used to detect the co-expression of proteins, and a Transwell assay and wound-healing assay were used to investigate the migration ability of cells. Experimental conclusions were verified in vivo using a nude mouse model. We first demonstrated overexpression of PAI-1 in gastric cancer tissues and cell lines. Subsequently, we found that NETs significantly enhanced the expression of EMT-related markers. These changes were accompanied by increases in cell invasion, migration, proliferation and tumour sphere formation. To further elucidate the mechanism, we employed lentivirus-mediated SERPINE-1 knockdown to reverse NET-induced EMT phenotype effectively. Mechanistically, we found that NETs activated the transforming growth factor (TGF)-β signalling pathway via PAI-1 as evidenced by increased expression of TGF-β1, TGF-βR1, TGF-βR2, phosphorylated Smad2/3 and Smad4. Finally, in vivo experiments using a nude mouse model of gastric cancer liver metastasis confirmed that NET-treated HGC-27 cells exhibited enhanced metastatic potential and SERPINE-1 knockdown abrogated metastatic potential. Our findings reveal a novel mechanism by which NETs promote EMT and metastasis in gastric cancer via the PAI-1–TGF-β axis. PAI-1 can be used as a potential target for the treatment of gastric cancer, and the expression of PAI-1 is closely related to the prognosis of patients with gastric cancer. Therapeutic strategies targeting NETs or PAI-1 may help prevent EMT and metastasis of gastric cancer and improve clinical outcomes in patients.
期刊介绍：
The Journal of Biochemical and Molecular Toxicology is an international journal that contains original research papers, rapid communications, mini-reviews, and book reviews, all focusing on the molecular mechanisms of action and detoxication of exogenous and endogenous chemicals and toxic agents. The scope includes effects on the organism at all stages of development, on organ systems, tissues, and cells as well as on enzymes, receptors, hormones, and genes. The biochemical and molecular aspects of uptake, transport, storage, excretion, lactivation and detoxication of drugs, agricultural, industrial and environmental chemicals, natural products and food additives are all subjects suitable for publication. Of particular interest are aspects of molecular biology related to biochemical toxicology. These include studies of the expression of genes related to detoxication and activation enzymes, toxicants with modes of action involving effects on nucleic acids, gene expression and protein synthesis, and the toxicity of products derived from biotechnology.
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