中国病理生理杂志2025，Vol.41Issue(5)：882-894,13.DOI:10.3969/j.issn.1000-4718.2025.05.006
石蒜碱通过YAP1/TRFC通路促进骨肉瘤细胞铁死亡并增强顺铂敏感性
Lycorine promotes ferroptosis in osteosarcoma cells via YAP1/TRFC path-way and enhances sensitivity to cisplatin
摘要
Abstract
AIM:This study aims to investigate the sensitizing effects of lycorine(Lyc)in combination with cisplatin(Cis)on human osteosarcoma cells and to explore the underlying mechanisms of action.METHODS:Human osteosarcoma cell lines MG63,HOS,and cisplatin-resistant HOS/DDP cells were utilized to evaluate the effects of Lyc and cisplatin,both alone and in combination,on cell viability using the CCK8 assay.The clonogenic assay was performed to assess cell proliferation capacity,while the scratch assay evaluated the drugs' effects on cell migration.Reactive oxygen species(ROS)levels were measured using a ROS assay kit,and changes in intracellular glutathione(GSH)levels were assessed with a GSH/oxidized glutathione(GSSG)assay kit.The mitochondrial membrane potential was analyzed via JC-1 staining to determine the drugs' effects on mitochondrial function.Intracellular iron(Ⅱ)content changes were detected us-ing FerrOrange,a fluorescence probe,and cellular malondialdehyde(MDA)levels were measured using an MDA assay kit.RT-qPCR was employed to evaluate the expression levels of key genes related to ferroptosis,and Western blot analysis was conducted to detect changes in the protein expression levels of Yes-associated protein 1(YAP1),acyl-CoA synthetase long-chain family member 4(ACSL4),glutathione peroxidase 4(GPX4),heme oxygenase-1(HO-1),and transferrin re-ceptor(TFRC).RESULTS:Both Lyc and cisplatin effectively inhibited the proliferation of human osteosarcoma cells.Notably,the combination of Lyc and cisplatin led to a more substantial reduction in cell viability,proliferation,and migra-tion abilities in MG63,HOS,and HOS/DDP cells compared to cisplatin alone.Additionally,this combination significant-ly increased ROS levels while decreasing GSH content,indicating mitochondrial damage and elevated iron(Ⅱ)and MDA levels.RT-qPCR results revealed that the combination treatment more significantly downregulated ferroptosis-promoting genes and upregulated ferroptosis-inhibiting genes compared to cisplatin treatment alone(P<0.05).Western blot results showed a slight decrease in GPX4 protein expression following Lyc and cisplatin treatment,while expression levels of YAP1,TFRC,ACSL4,and HO-1 were significantly increased(P<0.05).CONCLUSION:Lyc enhances the sensitivi-ty of MG63,HOS,and HOS/DDP cells to cisplatin by promoting ferroptosis through the YAP1/TFRC signaling pathway.关键词
石蒜碱/骨肉瘤/顺铂/耐药性/铁死亡Key words
lycorine/osteosarcoma/cisplatin/drug resistance/ferroptosis分类
临床医学引用本文复制引用
王林宽,王新平,易开君,黎智勇,赵琦,石义华..石蒜碱通过YAP1/TRFC通路促进骨肉瘤细胞铁死亡并增强顺铂敏感性[J].中国病理生理杂志,2025,41(5):882-894,13.基金项目
国家自然科学基金资助项目(No.81703015) （No.81703015）
湖北省自然科学基金创新发展联合基金资助项目(No.2024AFD059 （）
No.2023AFD042) （）
湖北省自然科学基金资助项目(No.2023AFB936) （No.2023AFB936）
湖北省卫生健康委科研项目(No.WJ2023F075) （No.WJ2023F075）
