Abstract：
BACKGROUND: Hepatocellular carcinoma (HCC) is a common malignancy, and ferroptosis is a novel form of cell death driven by excessive lipid peroxidation. In recent years, ferroptosis has been widely utilized in cancer treatment, and the ubiquitination modification system has been recognized to play a crucial role in tumorigenesis and metastasis. Increasing evidence suggests that ubiquitin regulates ferroptosis-related substrates involved in this process. However, the precise mechanism of utilizing ubiquitination modification to regulate ferroptosis for HCC treatment remains unclear.
METHODS: In this study, we detected the expression of TRIM33 in HCC using immunohistochemistry and western blotting techniques. The functional role of TRIM33 was verified through both in vitro and in vivo experiments. To evaluate the level of ferroptosis, mitochondrial superoxide levels, MDA levels, Fe2+ levels, and cell viability were assessed. Downstream substrates of TRIM33 were screened and confirmed via immunoprecipitation, immunofluorescence staining, and ubiquitination modification experiments.
RESULTS: Our findings demonstrate that TRIM33 inhibits the growth and metastasis of HCC cells both in vitro and in vivo while promoting their susceptibility to ferroptosis. Mechanistically speaking, TRIM33 induces cellular ferroptosis through E3 ligase-dependent degradation of TFRC-a known inhibitor of this process-thus elucidating the specific type and site at which TFRC undergoes modification by TRIM33.
CONCLUSIONS: In summary, our study reveals an important role for TRIM33 in HCC treatment while providing mechanistic support for its function. Additionally highlighted is the significance of ubiquitination modification leading to TFRC degradation-an insight that may prove valuable for future targeted therapies.
METHODS: In this study, we detected the expression of TRIM33 in HCC using immunohistochemistry and western blotting techniques. The functional role of TRIM33 was verified through both in vitro and in vivo experiments. To evaluate the level of ferroptosis, mitochondrial superoxide levels, MDA levels, Fe2+ levels, and cell viability were assessed. Downstream substrates of TRIM33 were screened and confirmed via immunoprecipitation, immunofluorescence staining, and ubiquitination modification experiments.
RESULTS: Our findings demonstrate that TRIM33 inhibits the growth and metastasis of HCC cells both in vitro and in vivo while promoting their susceptibility to ferroptosis. Mechanistically speaking, TRIM33 induces cellular ferroptosis through E3 ligase-dependent degradation of TFRC-a known inhibitor of this process-thus elucidating the specific type and site at which TFRC undergoes modification by TRIM33.
CONCLUSIONS: In summary, our study reveals an important role for TRIM33 in HCC treatment while providing mechanistic support for its function. Additionally highlighted is the significance of ubiquitination modification leading to TFRC degradation-an insight that may prove valuable for future targeted therapies.
摘要：
背景：肝细胞癌（HCC）是一种常见的恶性肿瘤，铁凋亡是一种由过度脂质过氧化引起的细胞死亡的新形式。近年来,在癌症治疗中,泛素化修饰系统在肿瘤发生和转移中起着至关重要的作用。越来越多的证据表明泛素调节参与该过程的铁凋亡相关底物。然而,利用泛素化修饰调节铁凋亡治疗HCC的确切机制尚不清楚.
方法：在本研究中，我们使用免疫组织化学和蛋白质印迹技术检测了TRIM33在HCC中的表达。通过体外和体内实验验证了TRIM33的功能作用。为了评估铁中毒的水平，线粒体超氧化物水平，MDA水平,Fe2+水平,并评估细胞活力。通过免疫沉淀筛选和确认TRIM33的下游底物，免疫荧光染色，和泛素化修饰实验。
结果：我们的发现表明，TRIM33在体外和体内抑制HCC细胞的生长和转移，同时促进其对铁凋亡的敏感性。机械地讲,TRIM33通过E3连接酶依赖性降解TFRC（该过程的已知抑制剂）诱导细胞铁凋亡，从而阐明了TFRC经过TRIM33修饰的特定类型和位点。
结论：总之，我们的研究揭示了TRIM33在HCC治疗中的重要作用,同时为其功能提供了机制支持.此外，还强调了泛素化修饰导致TFRC降解的重要性，这一见解可能对未来的靶向治疗有价值。
方法：在本研究中，我们使用免疫组织化学和蛋白质印迹技术检测了TRIM33在HCC中的表达。通过体外和体内实验验证了TRIM33的功能作用。为了评估铁中毒的水平，线粒体超氧化物水平，MDA水平,Fe2+水平,并评估细胞活力。通过免疫沉淀筛选和确认TRIM33的下游底物，免疫荧光染色，和泛素化修饰实验。
结果：我们的发现表明，TRIM33在体外和体内抑制HCC细胞的生长和转移，同时促进其对铁凋亡的敏感性。机械地讲,TRIM33通过E3连接酶依赖性降解TFRC（该过程的已知抑制剂）诱导细胞铁凋亡，从而阐明了TFRC经过TRIM33修饰的特定类型和位点。
结论：总之，我们的研究揭示了TRIM33在HCC治疗中的重要作用,同时为其功能提供了机制支持.此外，还强调了泛素化修饰导致TFRC降解的重要性，这一见解可能对未来的靶向治疗有价值。
