PMID- 38349263 OWN - NLM STAT- MEDLINE DCOM- 20240214 LR - 20240214 IS - 1098-2299 (Electronic) IS - 0272-4391 (Linking) VI - 85 IP - 1 DP - 2024 Feb TI - Everolimus inhibits hepatoblastoma by inducing autophagy-dependent ferroptosis. PG - e22140 LID - 10.1002/ddr.22140 [doi] AB - Everolimus, a known inhibitor of the mammalian target of rapamycin (mTOR), has shown uncertain efficacy in treating hepatoblastoma. This study delves into the potential anti-hepatoblastoma properties of everolimus and its intricate relationship with autophagy and ferroptosis, both in vitro and in vivo. In vivo, tumor tissue from hepatoblastoma patient and human hepatoblastoma cell line HuH-6 were xenografted into nude mice to establish xenograft models for observing the effect of everolimus on tumor growth. In vitro, HuH-6 cells were cultured to evaluate the anti-hepatoblastoma activity of everolimus. Transmission electron microscopy and microtubule-associated proteins 1 light chain 3 (LC3), beclin 1, and p62 protein expressions were employed to investigate autophagy. Additionally, indicators of cell apoptosis, reactive oxygen species (ROS) and proteins associated with ferroptosis were measured to evaluate ferroptosis. The results demonstrate that everolimus treatment effectively induced the formation of autophagosomes in hepatoblastoma cells, upregulated the LC3II/I ratio and beclin 1 expression, and downregulated p62 expression, indicating an enhanced autophagy level both in vitro and in vivo. Furthermore, everolimus treatment induced cell apoptosis, increased ROS level, elevated concentrations of malondialdehyde, 4-hydroxynonenal, and iron content, while reducing the ratio of glutathione/oxidized glutathione, and downregulating the protein expression of glutathione peroxidase 4 and solute carrier family 7 member 11, suggesting its ability to induce ferroptosis in hepatoblastoma cells. Importantly, the induction of ferroptosis by everolimus was significantly reversed in the presence of autophinib, an autophagy inhibitor, indicating the autophagy-dependent of everolimus-induced ferroptosis. Taken together, these findings suggest that everolimus holds promise as an effective anti-hepatoblastoma drug, with its mechanism of action potentially involving the induction of autophagy-dependent ferroptosis in hepatoblastoma cells. CI - (c) 2023 Wiley Periodicals LLC. FAU - Huang, Haijin AU - Huang H AD - Department of Pediatric Surgery, First Affiliated Hospital of Gannan Medical University, Ganzhou, Jiangxi, China. AD - Jiangxi Provincial Clinical Research Center for Vascular Anomalies, The First Affiliated Hospital of GanNan Medical University, Ganzhou, Jiangxi, China. FAU - Yan, Jinlong AU - Yan J AUID- ORCID: 0000-0002-7040-0275 AD - Department of General Surgery, Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, China. FAU - Xu, Xianyun AU - Xu X AD - Department of Clinical Laboratory, Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine, Nanchang, Jiangxi, China. FAU - Feng, Yanping AU - Feng Y AD - Department of Neurological Surgery, First Affiliated Hospital of Gannan Medical University, Ganzhou, Jiangxi, China. FAU - Liu, Haijin AU - Liu H AD - Department of Pediatric Surgery, First Affiliated Hospital of Gannan Medical University, Ganzhou, Jiangxi, China. FAU - Liu, Jianping AU - Liu J AD - Department of General Surgery, First Affiliated Hospital of Gannan Medical University, Ganzhou, Jiangxi, China. FAU - Xie, Mingfeng AU - Xie M AD - Jiangxi Provincial Clinical Research Center for Vascular Anomalies, The First Affiliated Hospital of GanNan Medical University, Ganzhou, Jiangxi, China. AD - Chinese & Western Integrative Medicine Discipline, Jiangxi University of Chinese Medicine, Nanchang, Jiangxi, China. AD - Jiangxi Key Laboratory of TCM for Prevention and Treatment on Hemangioma, Nanchang, Jiangxi, China. AD - Integrated Chinese and Western Medicine Institute for Children Health & Drug Innovation, Jiangxi University of Chinese Medicine, Nanchang, Jiangxi, China. FAU - Chen, Leifeng AU - Chen L AD - Department of General Surgery, Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi, China. FAU - Xiang, Deng AU - Xiang D AD - Department of General Surgery, The Affiliated Children's Hospital of Nanchang Medical College, Nanchang, Jiangxi, China. FAU - Peng, Wei AU - Peng W AD - Department of Pediatric Surgery, First Affiliated Hospital of Gannan Medical University, Ganzhou, Jiangxi, China. FAU - Zeng, Linshan AU - Zeng L AD - Department of Pediatric Surgery, First Affiliated Hospital of Gannan Medical University, Ganzhou, Jiangxi, China. FAU - Zeng, Yong AU - Zeng Y AD - Department of Pediatric Surgery, First Affiliated Hospital of Gannan Medical University, Ganzhou, Jiangxi, China. FAU - Chen, Feng AU - Chen F AD - Department of Pediatric Surgery, First Affiliated Hospital of Gannan Medical University, Ganzhou, Jiangxi, China. FAU - Zhang, Shouhua AU - Zhang S AD - Department of General Surgery, The Affiliated Children's Hospital of Nanchang Medical College, Nanchang, Jiangxi, China. FAU - Liu, Qian AU - Liu Q AUID- ORCID: 0009-0003-3564-9635 AD - Jiangxi Provincial Clinical Research Center for Vascular Anomalies, The First Affiliated Hospital of GanNan Medical University, Ganzhou, Jiangxi, China. AD - Chinese & Western Integrative Medicine Discipline, Jiangxi University of Chinese Medicine, Nanchang, Jiangxi, China. AD - Jiangxi Key Laboratory of TCM for Prevention and Treatment on Hemangioma, Nanchang, Jiangxi, China. AD - Integrated Chinese and Western Medicine Institute for Children Health & Drug Innovation, Jiangxi University of Chinese Medicine, Nanchang, Jiangxi, China. LA - eng GR - 20181BAB205052/Nature Science Foundation of Jiangxi/ GR - 20192ACBL20005/Nature Science Foundation of Jiangxi/ GR - XN202026/Key Laboratory of Cardiovascular and Cerebrovascular Diseases Prevention and Treatment of Ministry of Education (Rare Earth Special Project)/ GR - GJJ2201454/Jiangxi Province Office of Education Support Program/ GR - GZ2023ZSF112/Ganzhou Guiding Science and Technology Project/ GR - 82260570/National Natural Science Foundation of China/ GR - 202130879/Science and Technology Plan Project of Jiangxi Provincial Health Commission/ PT - Journal Article PL - United States TA - Drug Dev Res JT - Drug development research JID - 8204468 RN - 9HW64Q8G6G (Everolimus) RN - 0 (Beclin-1) RN - 0 (Reactive Oxygen Species) SB - IM MH - Animals MH - Mice MH - Humans MH - Everolimus/pharmacology MH - *Hepatoblastoma/drug therapy MH - Beclin-1 MH - *Ferroptosis MH - Mice, Nude MH - Reactive Oxygen Species MH - Autophagy MH - *Liver Neoplasms/drug therapy MH - Mammals OTO - NOTNLM OT - autophagy OT - everolimus OT - ferroptosis OT - hepatoblastoma EDAT- 2024/02/13 12:47 MHDA- 2024/02/13 12:48 CRDT- 2024/02/13 10:03 PHST- 2023/11/22 00:00 [revised] PHST- 2023/08/28 00:00 [received] PHST- 2023/12/08 00:00 [accepted] PHST- 2024/02/13 12:48 [medline] PHST- 2024/02/13 12:47 [pubmed] PHST- 2024/02/13 10:03 [entrez] AID - 10.1002/ddr.22140 [doi] PST - ppublish SO - Drug Dev Res. 2024 Feb;85(1):e22140. doi: 10.1002/ddr.22140.