Ponicidin triggered ferroptosis in esophageal squamous cell carcinoma by suppressing the SLC7A11/Glutathione/GPX4 signalling axis

IF 6.7 1区医学 Q1 CHEMISTRY, MEDICINAL

Phytomedicine Pub Date : 2025-05-28 DOI:10.1016/j.phymed.2025.156925

Wenhu Liu , Jinhua Zhang , Min Wu , Dan Ren , Chuan Chen , Zuo Du , Qianhui Li , Jinxia Chang , Qi Pu , Zhenzhong Liu

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引用次数: 0

Abstract

Background

Ponicidin, a diterpenoid derived from Rabdosia rubescens, exhibits potent antitumor activity. However, its mechanisms against esophageal squamous cell carcinoma (ESCC) remain obscure. This study aims to explore the effects of ponicidin against ESCC and reveal its molecular mechanisms.

Methods

The anti-ESCC effects of ponicidin were evaluated using CCK-8 assay, colony formation and transwell invasion assays. Cell cycle progression and mitochondrial membrane potential were analyzed using flow cytometry. Proteomics was applied to explore ponicidin’s mechanisms. Ferroptosis induction was evaluated by quantifying reactive oxygen species, Fe²⁺, malondialdehyde, glutathione, and lipid peroxidation levels. Docking and molecular dynamics simulations were conducted to identify the targets. siRNA was employed to validate target. The efficacy of ponicidin on tumorigenicity was explored in tumor xenograft mouse models, and its biosafety was evaluated via hemolysis assays, plasma ALT, AST, BUN, and CRE levels, as well as histopathological examinations. Western blot was used to analyze protein expression levels.

Results

Ponicidin inhibited ESCC cell proliferation, arrested cells in the G₂/M phase, reduced mitochondrial membrane potential, and suppressed tumor growth without evident toxicity. Proteomics identified that ponicidin-induced ferroptosis is the predominant mechanism against ESCC. Ponicidin increased reactive oxygen species, malondialdehyde, Fe²⁺, lipid peroxidation and glutathione depletion. Ferrostatin-1 pretreatment reduced lipid peroxidation, rescued PON induced inhibition of cell viability, and reversed the decreased expression of SLC7A11, GPX4 and GSR. Molecular docking revealed strong binding affinity of PON to GPX4 (-7.31±0.55 kcal/mol) and SLC7A11 (-8.19±0.37 kcal/mol). Molecular dynamics simulations confirmed stabilized complexes with total interaction energies of -23.43 ± 2.13 kcal/mol (GPX4-PON) and -31.42 ± 0.84 kcal/mol (SLC7A11-PON). siRNA-mediated knockdown of GPX4 and SLC7A11 reduced ESCC sensitivity to ponicidin-induced ferroptosis.

Conclusion

This study provides the first evidence that ponicidin triggers ferroptosis in ESCC cells via suppression of the SLC7A11/glutathione/GPX4 signalling axis, offering actionable targets for ferroptosis-enhancing combination therapies.

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Ponicidin通过抑制SLC7A11/谷胱甘肽/GPX4信号轴引发食管鳞状细胞癌的铁下垂

研究背景：从冬凌草中提取的二萜类化合物ponicidin具有很强的抗肿瘤活性。然而，其抗食管鳞状细胞癌（ESCC）的机制尚不清楚。本研究旨在探讨poniciidin对ESCC的作用及其分子机制。方法采用CCK-8法、菌落形成法和跨井侵袭法评价ponicidin的抗escc作用。流式细胞术分析细胞周期进程和线粒体膜电位。利用蛋白质组学方法探讨了ponicidin的作用机制。通过量化活性氧、Fe2+、丙二醛、谷胱甘肽和脂质过氧化水平来评估铁下垂诱导。通过对接和分子动力学模拟来识别目标。采用siRNA对靶标进行验证。在肿瘤异种移植小鼠模型中探讨了ponicidin的致瘤性，并通过溶血试验、血浆ALT、AST、BUN和CRE水平以及组织病理学检查来评价其生物安全性。Western blot检测蛋白表达水平。结果sponciidin抑制ESCC细胞增殖，使细胞停留在G2/M期，降低线粒体膜电位，抑制肿瘤生长，但无明显毒性。蛋白质组学发现，poniciidin诱导的铁下垂是ESCC的主要机制。poniciidin增加活性氧、丙二醛、Fe2+、脂质过氧化和谷胱甘肽消耗。铁抑素-1预处理降低了脂质过氧化，挽救了PON诱导的细胞活力抑制，逆转了SLC7A11、GPX4和GSR的表达下降。分子对接显示PON与GPX4（-7.31±0.55 kcal/mol）和SLC7A11（-8.19±0.37 kcal/mol）具有较强的结合亲和力。分子动力学模拟证实，配合物的总相互作用能分别为-23.43±2.13 kcal/mol （GPX4-PON）和-31.42±0.84 kcal/mol （SLC7A11-PON）。sirna介导的敲低GPX4和SLC7A11可降低ESCC对猪毒素诱导的铁下垂的敏感性。结论本研究首次证明了ponicidin通过抑制SLC7A11/谷胱甘肽/GPX4信号轴触发ESCC细胞铁凋亡，为铁凋亡增强联合治疗提供了可操作的靶点。

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

Phytomedicine 医学-药学

CiteScore

10.30

自引率

5.10%

发文量

670

审稿时长

91 days

期刊介绍： Phytomedicine is a therapy-oriented journal that publishes innovative studies on the efficacy, safety, quality, and mechanisms of action of specified plant extracts, phytopharmaceuticals, and their isolated constituents. This includes clinical, pharmacological, pharmacokinetic, and toxicological studies of herbal medicinal products, preparations, and purified compounds with defined and consistent quality, ensuring reproducible pharmacological activity. Founded in 1994, Phytomedicine aims to focus and stimulate research in this field and establish internationally accepted scientific standards for pharmacological studies, proof of clinical efficacy, and safety of phytomedicines.