Plumbagin activates NLRP3/caspase-1/GSDMD-mediated pyroptosis by decreasing DUSP4 expression to exhibit anticancer property in cholangiocarcinoma

IF 3.4 3区医学 Q2 PHARMACOLOGY & PHARMACY

Toxicology and applied pharmacology Pub Date : 2025-07-29 DOI:10.1016/j.taap.2025.117492

Fei Song , Lingxiao Wei , Junchao Wu , Guoyi Zhu , Jigui Zhang , Hongyang Li

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引用次数: 0

Abstract

Background

Plumbagin is effective in treating cholangiocarcinoma (CCA), however, the potential mechanism of plumbagin therapy is still unclear. This study aims to uncover the anti-tumor nature of plumbagin and the specific mechanism by which plumbagin alleviates NLRP3/caspase-1/GSDMD-induced pyroptosis.

Methods

Human CCA cells lines HuCCT1 and RBE and the CCA mouse model injected with HuCCT1 cells were used for the mechanism exploration. Different concentrations of plumbagin were incubated with the cells or injected to the CCA mouse to investigate the anti-tumor role of pumbagin. CCK-8 and Transwell was used to detect the cell viability and migration, respectively. ELISA kits were used to detect the ROS, LDH, IL-1β, and IL-18 levels. SEM was used for pyroptosis condition observation. qRT-PCR and WB were used to detect the expression levels of DUSP4, NLRP3, GSDMD-N, and cle-caspase-1 levels. Immunohistochemical staining was used for detection of tumor tissue injury.

Results

In CCA cells, plumbagin inhibited the cell viability and migration. In CCA mouse model, plumbagin inhibited the tumor growth. Along with the plumbagin treatment, the decreased DUSP4 expression, and increased ROS, LDH, NLRP3, clecaspase-1, GSDMD-N proteins, and pyroptosis degree, were investigated. After addition of NLRP3 inhibitor MCC950 and transfection of oe-DUSP4, the promoted pyroptosis and inhibited tumor growth induced by plumbagin were reversed, indicating that the plumbagin treatment inhibited the CCA progression through decreasing the DUSP4 expression and thus activating the pyroptosis.

Conclusion

DUSP4/NLRP3/caspase-1/GSDMD-mediated pyroptosis is demonstrated to be the therapeutic mechanism of plumbagin in CCA.

Abstract Image

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白丹素通过降低DUSP4表达激活NLRP3/caspase-1/ gsdmd介导的焦亡，在胆管癌中表现出抗癌特性。

背景：白桦素治疗胆管癌（CCA）有效，但其治疗的潜在机制尚不清楚。本研究旨在揭示白桦苷的抗肿瘤性质，以及白桦苷缓解NLRP3/caspase-1/ gsdmd诱导的焦亡的具体机制。方法：采用人CCA细胞系HuCCT1和RBE，以及注射HuCCT1细胞的CCA小鼠模型进行机制探索。以不同浓度的白桦素与细胞孵育或注射于CCA小鼠体内，研究白桦素的抗肿瘤作用。CCK-8和Transwell分别检测细胞活力和迁移率。采用ELISA试剂盒检测ROS、LDH、IL-1β、IL-18水平。用扫描电镜观察焦亡情况。采用qRT-PCR和WB检测DUSP4、NLRP3、GSDMD-N和cle-caspase-1的表达水平。采用免疫组化染色检测肿瘤组织损伤。结果：在CCA细胞中，白桦素抑制细胞活力和迁移。在CCA小鼠模型中，白桦素抑制肿瘤生长。随着白杨桃素的处理，DUSP4的表达降低，ROS、LDH、NLRP3、clecaspase-1、GSDMD-N蛋白的表达升高，焦亡程度升高。添加NLRP3抑制剂MCC950并转染e-DUSP4后，白桦素诱导的促焦亡和抑制肿瘤生长的作用发生逆转，说明白桦素通过降低DUSP4的表达从而激活焦亡来抑制CCA的进展。结论：DUSP4/NLRP3/caspase-1/ gsdmd介导的焦亡可能是白杨苷治疗CCA的机制。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Toxicology and applied pharmacology 医学-毒理学

CiteScore

6.80

自引率

2.60%

发文量

309

审稿时长

32 days

期刊介绍： Toxicology and Applied Pharmacology publishes original scientific research of relevance to animals or humans pertaining to the action of chemicals, drugs, or chemically-defined natural products. Regular articles address mechanistic approaches to physiological, pharmacologic, biochemical, cellular, or molecular understanding of toxicologic/pathologic lesions and to methods used to describe these responses. Safety Science articles address outstanding state-of-the-art preclinical and human translational characterization of drug and chemical safety employing cutting-edge science. Highly significant Regulatory Safety Science articles will also be considered in this category. Papers concerned with alternatives to the use of experimental animals are encouraged. Short articles report on high impact studies of broad interest to readers of TAAP that would benefit from rapid publication. These articles should contain no more than a combined total of four figures and tables. Authors should include in their cover letter the justification for consideration of their manuscript as a short article.