Hou Chun-Yu, Suo Yu-Hong, Lv Pan, Yuan Hong-Feng, Zhao Li-Na, Wang Yu-Fei, Zhang Hui-Hui, Sun Jiao, Sun Lin-Lin, Lu Wei, Zhang Ning-Ning, Yang Guang, Zhang Xiao-Dong
National Key Laboratory of Draggability Evaluation and Systematic Translational Medicine, Tianjin's Clinical Research Center for Cancer, Tianjin Key Laboratory of Digestive Cancer, Department of Gastrointestinal Cancer Biology, Tianjin Cancer Institute, Liver Cancer Center, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin, 300060, China.
Department of Hepatobiliary Oncology, Liver Cancer Center, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin, 300060, China.
Acta Pharmacol Sin. 2025 Jan;46(1):208-221. doi: 10.1038/s41401-024-01354-0. Epub 2024 Aug 1.
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.
马兜铃酸（AAs）已被确认为肝细胞癌（HCC）的一个重要风险因素。铁死亡是一种参与肿瘤发展的程序性细胞死亡类型。在本研究中，我们探究了AAs促进HCC生长的分子机制。通过生物信息学和RNA测序分析，我们发现AAs与铁死亡密切相关。通过生物信息学分析以及表面等离子体共振（SPR）分析，利用包含p53的Pro92、Arg174、Asp207、Phe212和His214的结合口袋位点，验证了HepG2细胞中p53与AAs之间的物理相互作用。基于与AAs相互作用的结合口袋，我们设计了一个突变体并进行了RNA测序分析。有趣的是，我们发现该结合口袋与铁死亡、生长停滞和DNA损伤诱导蛋白45α（GADD45A）、核因子E2相关因子2（NRF2）以及溶质载体家族7成员11（SLC7A11）有关。在功能上，这种相互作用干扰了p53与GADD45A或NRF2启动子的结合，削弱了p53在增强GADD45A和抑制NRF2方面的作用；而突变体并未表现出相同的效果。因此，这一事件下调了GADD45A并上调了NRF2，最终抑制了铁死亡，表明在HepG2细胞中AAs劫持了p53来下调GADD45A并上调NRF2。因此，AAs处理通过p53/GADD45A/NRF2/SLC7A11轴导致铁死亡受到抑制，进而导致肿瘤生长增强。总之，AAs劫持的p53通过GADD45A/NRF2/SLC7A11轴抑制铁死亡以增强肿瘤生长。我们的研究结果提供了AAs促进HCC的潜在机制以及对肝癌中p53的新见解。在治疗方面，癌基因NRF2是肝癌一个有前景的靶点。
