【作者】 陈浩；

【导师】 于光涛；

【作者基本信息】 南方医科大学 ， 口腔颌面外科学（专业学位）， 2025， 硕士

【摘要】 研究背景及目的口腔鳞状细胞癌为头颈部肿瘤最常见的病理类型之一。然而,现有的治疗策略无法显著改善口腔鳞状细胞癌患者的预后。铁死亡被定义为一种新型的调节性细胞死亡,其特征是铁依赖性脂质过氧化。近年来,铁死亡在癌症治疗中的研究取得了实质性进展。有许多与铁死亡触发相关的酶已经得到鉴定,其中最突出的是长链酰基辅酶A合成酶4(ACSL4)。课题组前期研究证实了锰基纳米材料能够引起肿瘤细胞的铁死亡,但具体机制不明。本研究的目的是探讨锰离子是否能诱导口腔鳞状细胞癌的铁死亡及其潜在的生物学机制。材料和方法用CCK8评价锰对口腔鳞癌细胞的毒性作用。然后,运用RNA测序法筛选锰处理组差异表达的铁死亡相关基因。通过蛋白印迹及酶联免疫吸附测定法(ELISA)确定YAP/TAZ-ACSL4铁死亡信号通路的激活。免疫荧光显微术解析YAP/TAZ相分离特征。体内实验构建BALB/c-nu小鼠口腔鳞癌移植瘤模型验证锰驱动铁死亡的体内抗肿瘤效应。最后,应用口腔鳞状细胞癌细胞患者临床标本芯片分析YAP/TAZ-ACSL4的临床意义。结果CCK8结果显示锰处理的口腔鳞癌细胞活性被显著抑制。RNA测序分析结果表明锰处理后铁死亡关键基因ACSL4和YAP/TAZ上调。ELISA、蛋白印迹等结果进一步证实,锰诱导的铁死亡依赖于YAP/TAZ-ACSL4信号通路。此外,通过免疫荧光实验证实了 ACSL4的活化是通过YAP/TAZ相分离实现的。体内实验证实锰作为铁死亡触发器显著抑制了口腔鳞癌的生长。临床数据分析表明,YAP/TAZ-ACSL4轴表达水平与患者生存期呈正相关。结论本研究首次阐明锰通过调控YAP/TAZ相分离激活ACSL4的铁死亡信号通路,为口腔鳞癌治疗提供了新型治疗策略。临床数据证实YAP/TAZ-ACSL4轴可作为预后评估的分子标志物,为开发锰基铁死亡诱导剂奠定了理论基础。更多还原

【Abstract】 BackgroundOral squamous cell carcinoma(OSCC)is one of the most common pathological types of head and neck tumors.However,existing treatment strategies cannot significantly improve the prognosis of patients with OSCC.Ferroptosis is defined as a novel regulatory cell death characterized by iron dependent lipid peroxidation.In recent years,the role of ferroptosis in cancer treatment has been confirmed and substantial progress has been made.Many enzymes related to ferroptosis triggering have been identified,among which the most prominent is long-chain acyl CoA synthetase 4(ACSL4).Our Previous research has confirmed that manganese-based nanomaterials can induce ferroptosis in tumor cells,but the specific mechanism is unclear.The purpose of this study is to investigate whether manganese can induce ferroptosis in oral squamous cell carcinoma and its potential biological mechanisms.Materials and MethodsThe cytotoxic effects of manganese on OSCC were quantitatively assessed using Cell Counting Kit-8(CCK8)assays.RNA sequencing was subsequently performed to identify ferroptosis-associated genes exhibiting differential expression in manganesetreated groups.Activation of the YAP/TAZ-ACSL4 ferroptosis signaling axis was systematically verified through Western blot analysis and enzyme-linked immunosorbent assay(ELISA).The liquid-liquid phase separation(LLPS)characteristics of YAP/TAZ complexes were elucidated via immunofluorescence microscopy.In vivo antitumor efficacy of manganese-induced ferroptosis was validated using orthotopic OSCC xenograft models in BALB/c nude mice.Clinical relevance was investigated through tissue microarray analysis of YAP/TAZ/ACSL4 expression patterns in human OSCC specimens.ResultsThe CCK8 results showed that the activity of OSCC cells treated with manganese was significantly inhibited.RNA sequencing revealed significant upregulation of ferroptosis-related genes ACSL4 and YAP/TAZ following manganese exposure.Multimodal validation through ELISA and Western blotting confirmed manganese-induced ferroptosis mechanistically depends on YAP/TAZ-ACSL4 signaling transduction.Phase separation dynamics analysis demonstrated YAP/TAZ LLPS mediates ACSL4 activation.In vivo experiments substantiated manganese’s tumor-suppressive effects through ferroptosis induction in xenograft models.Clinical correlation analysis established positive associations between YAP/TAZ-ACSL4 axis expression levels and patient survival outcomes.ConclusionThis study elucidates for the first time that manganese activates the ACSL4-mediated ferroptosis pathway by regulating YAP/TAZ phase separation,unveiling novel therapeutic avenues for OSCC management.Clinical validation identifies the YAP/TAZ-ACSL4 axis as a potential prognostic biomarker,establishing theoretical foundations for developing manganese-based ferroptosis-inducing therapeutics.更多还原