许多研究表明，阿司匹林作为常用药物，可以预防肝细胞癌（HCC）的发展。铁死亡作为一种动态的肿瘤抑制剂，在肝癌发生中发挥着重要作用。在本研究中，我们调查了阿司匹林是否影响肝癌细胞中的铁死亡。RNA-seq分析表明，在经过阿司匹林处理的HepG2细胞中，阿司匹林上调了4个与铁死亡相关的驱动基因，并下调了5个与铁死亡相关的抑制基因。使用阿司匹林（4mM）的治疗诱导了HepG2和Huh7细胞中的铁死亡，而这一作用可以通过铁死亡诱导剂恶嗪（10μM）增强。我们证明，NF-κB p65通过直接结合SLC7A11启动子的核心区域并激活铁死亡抑制剂SLC7A11的转录来限制HepG2和Huh7细胞的铁死亡，而阿司匹林则通过抑制NF-κB p65激活的SLC7A11转录来诱导铁死亡。p65的过表达可以拯救HepG2和Huh7细胞免于阿司匹林诱导的铁死亡。高表达SLC7A11和p65的HCC患者存活率较低。在体内外，NF-κB p65功能阻止了阿司匹林诱导的铁死亡，而恶嗪可以减弱这种阻止作用。我们得出结论，阿司匹林限制NF-κB激活的SLC7A11转录以诱导铁死亡来抑制HCC的生长。这些结果为阿司匹林调节肝癌中铁死亡的机制提供了新的见解。阿司匹林和铁死亡诱导剂的联合使用可能为HCC的治疗提供一种潜在策略。©2023作者，独家授权给中国科学院上海药物研究所和中国药理学会。

A number of studies have shown that aspirin, as commonly prescribed drug, prevents the development of hepatocellular carcinoma (HCC). Ferroptosis as a dynamic tumor suppressor plays a vital role in hepatocarcinogenesis. In this study we investigated whether aspirin affected ferroptosis in liver cancer cells. RNA-seq analysis revealed that aspirin up-regulated 4 ferroptosis-related drivers and down-regulated 5 ferroptosis-related suppressors in aspirin-treated HepG2 cells. Treatment with aspirin (4 mM) induced remarkable ferroptosis in HepG2 and Huh7 cells, which was enhanced by the ferroptosis inducer erastin (10 μM). We demonstrated that NF-κB p65 restricted ferroptosis in HepG2 and Huh7 cells through directly binding to the core region of SLC7A11 promoter and activating the transcription of ferroptosis inhibitor SLC7A11, whereas aspirin induced ferroptosis through inhibiting NF-κB p65-activated SLC7A11 transcription. Overexpression of p65 rescued HepG2 and Huh7 cells from aspirin-induced ferroptosis. HCC patients with high expression levels of SLC7A11 and p65 presented lower survival rate. Functionally, NF-κB p65 blocked the aspirin-induced ferroptosis in vitro and in vivo, which was attenuated by erastin. We conclude that aspirin triggers ferroptosis by restricting NF-κB-activated SLC7A11 transcription to suppress the growth of HCC. These results provide a new insight into the mechanism by which aspirin regulates ferroptosis in hepatocarcinogenesis. A combination of aspirin and ferroptosis inducer may provide a potential strategy for the treatment of HCC in clinic.© 2023. The Author(s), under exclusive licence to Shanghai Institute of Materia Medica, Chinese Academy of Sciences and Chinese Pharmacological Society.