马兜铃酸 (AA) 已被确定为肝细胞癌 (HCC) 的重要危险因素。铁死亡是一种参与肿瘤发展的受调节细胞死亡。在这项研究中，我们研究了AA促进HCC生长的分子机制。通过生物信息学和RNA-Seq分析，我们发现AA与铁死亡密切相关。通过生物信息学分析和 SPR 测定（使用包含 p53 的 Pro92、Arg174、Asp207、Phe212 和 His214 的结合袋位点）验证了 HepG2 细胞中 p53 和 AA 之间的物理相互作用。基于与 AA 相互作用的结合口袋，我们设计了一个突变体并进行了 RNA-Seq 分析。有趣的是，我们发现结合口袋导致铁死亡、GADD45A、NRF2 和 SLC7A11。从功能上讲，这种相互作用干扰了 p53 与 GADD45A 或 NRF2 启动子的结合，削弱了 p53 增强 GADD45A 和抑制 NRF2 的作用；突变体没有表现出相同的效果。因此，该事件下调了 GADD45A 并上调了 NRF2，最终抑制了铁死亡，表明 AA 劫持了 p53，从而下调了 HepG2 细胞中的 GADD45A 并上调了 NRF2。因此，AA 处理通过 p53/GADD45A/NRF2/SLC7A11 轴抑制铁死亡，从而促进肿瘤生长。总之，AAs 劫持的 p53 通过 GADD45A/NRF2/SLC7A11 轴抑制铁死亡，从而促进肿瘤生长。我们的研究结果提供了 AA 增强 HCC 的潜在机制，以及对肝癌中 p53 的新见解。在治疗上，癌基因 NRF2 是肝癌的一个有前景的靶标。© 2024。作者获得中国科学院上海药物研究所和中国药理学会的独家许可。

Aristolochic acids (AAs) have been identified as a significant risk factor for hepatocellular carcinoma (HCC). Ferroptosis is a type of regulated cell death involved in the tumor development. In this study, we investigated the molecular mechanisms by which AAs enhanced the growth of HCC. By conducting bioinformatics and RNA-Seq analyses, we found that AAs were closely correlated with ferroptosis. The physical interaction between p53 and AAs in HepG2 cells was validated by bioinformatics analysis and SPR assays with the binding pocket sites containing Pro92, Arg174, Asp207, Phe212, and His214 of p53. Based on the binding pocket that interacts with AAs, we designed a mutant and performed RNA-Seq profiling. Interestingly, we found that the binding pocket was responsible for ferroptosis, GADD45A, NRF2, and SLC7A11. Functionally, the interaction disturbed the binding of p53 to the promoter of GADD45A or NRF2, attenuating the role of p53 in enhancing GADD45A and suppressing NRF2; the mutant did not exhibit the same effects. Consequently, this event down-regulated GADD45A and up-regulated NRF2, ultimately inhibiting ferroptosis, suggesting that AAs hijacked p53 to down-regulate GADD45A and up-regulate NRF2 in HepG2 cells. Thus, AAs treatment resulted in the inhibition of ferroptosis via the p53/GADD45A/NRF2/SLC7A11 axis, which led to the enhancement of tumor growth. In conclusion, AAs-hijacked p53 restrains ferroptosis through the GADD45A/NRF2/SLC7A11 axis to enhance tumor growth. Our findings provide an underlying mechanism by which AAs enhance HCC and new insights into p53 in liver cancer. Therapeutically, the oncogene NRF2 is a promising target for liver cancer.© 2024. The Author(s), under exclusive licence to Shanghai Institute of Materia Medica, Chinese Academy of Sciences and Chinese Pharmacological Society.