PDF(Pubmed)
Abstract：
BACKGROUND: Hepatocellular carcinoma (HCC) ranks as the second leading cause of global cancer-related deaths and is characterized by a poor prognosis. Eukaryotic translation elongation factor 1 alpha 1 (EEF1A1) have been proved to play important roles in various human cancers, whereas the deubiquitination of EEF1A1 was poorly understood.
METHODS: The binding and regulatory relationship between Ubiquitin carboxyl-terminal hydrolase L3 (UCHL3) and EEF1A1 was validated using clinical tissue samples, reverse transcription quantitative real-time fluorescence quantitative PCR (RT-qPCR), Western blotting, co-immunoprecipitation, and immunofluorescence, as well as ubiquitin detection and cyclohexamide tracking experiments. Finally, the impact of the UCHL3/EEF1A1 axis on HCC malignant behavior was analyzed through functional experiments and nude mouse models.
RESULTS: UCHL3 was found to have a high expression level in HCC tissues. Tissue samples from 60 HCC patients were used to evaluate the correlation between UCHL3 and EEF1A1. UCHL3 binds to EEF1A1 through the lysine site, which reduces the ubiquitination level of EEF1A1. Functional experiments and nude mouse models have demonstrated that the UCHL3/EEF1A1 axis promotes the migration, stemness, and drug resistance of HCC cells. Reducing the expression of EEF1A1 can reverse the effect of UCHL3 on the malignant behavior of HCC cells.
CONCLUSIONS: Our findings revealed that UCHL3 binds and stabilizes EEF1A1 through deubiquitination. UCHL3 and EEF1A1 formed a functional axis in facilitating the malignant progression of HCC, proving new insights for the anti-tumor targeted therapy for HCC.
METHODS: The binding and regulatory relationship between Ubiquitin carboxyl-terminal hydrolase L3 (UCHL3) and EEF1A1 was validated using clinical tissue samples, reverse transcription quantitative real-time fluorescence quantitative PCR (RT-qPCR), Western blotting, co-immunoprecipitation, and immunofluorescence, as well as ubiquitin detection and cyclohexamide tracking experiments. Finally, the impact of the UCHL3/EEF1A1 axis on HCC malignant behavior was analyzed through functional experiments and nude mouse models.
RESULTS: UCHL3 was found to have a high expression level in HCC tissues. Tissue samples from 60 HCC patients were used to evaluate the correlation between UCHL3 and EEF1A1. UCHL3 binds to EEF1A1 through the lysine site, which reduces the ubiquitination level of EEF1A1. Functional experiments and nude mouse models have demonstrated that the UCHL3/EEF1A1 axis promotes the migration, stemness, and drug resistance of HCC cells. Reducing the expression of EEF1A1 can reverse the effect of UCHL3 on the malignant behavior of HCC cells.
CONCLUSIONS: Our findings revealed that UCHL3 binds and stabilizes EEF1A1 through deubiquitination. UCHL3 and EEF1A1 formed a functional axis in facilitating the malignant progression of HCC, proving new insights for the anti-tumor targeted therapy for HCC.
摘要：
背景：肝细胞癌（HCC）是全球癌症相关死亡的第二大原因，其特征是预后不良。真核翻译延伸因子1α1(EEF1A1)已被证明在多种人类癌症中发挥重要作用,而对EEF1A1的去泛素化知之甚少。
方法：使用临床组织样本验证了泛素羧基末端水解酶L3（UCHL3）与EEF1A1之间的结合和调节关系，逆转录定量实时荧光定量PCR(RT-qPCR),西方印迹,免疫共沉淀,和免疫荧光，以及泛素检测和环己酰胺跟踪实验。最后,通过功能实验和裸鼠模型分析了UCHL3/EEF1A1轴对HCC恶性行为的影响。
结果：发现UCHL3在HCC组织中具有高表达水平。来自60例HCC患者的组织样本用于评估UCHL3和EEF1A1之间的相关性。UCHL3通过赖氨酸位点与EEF1A1结合，这降低了EEF1A1的泛素化水平。功能实验和裸鼠模型已经证明UCHL3/EEF1A1轴促进迁移,stemness,和肝癌细胞的耐药性。降低EEF1A1的表达可以逆转UCHL3对HCC细胞恶性行为的影响。
结论：我们的发现表明UCHL3通过去泛素化结合并稳定EEF1A1。UCHL3和EEF1A1形成促进HCC恶性进展的功能轴，为肝癌的抗肿瘤靶向治疗提供新的见解。
方法：使用临床组织样本验证了泛素羧基末端水解酶L3（UCHL3）与EEF1A1之间的结合和调节关系，逆转录定量实时荧光定量PCR(RT-qPCR),西方印迹,免疫共沉淀,和免疫荧光，以及泛素检测和环己酰胺跟踪实验。最后,通过功能实验和裸鼠模型分析了UCHL3/EEF1A1轴对HCC恶性行为的影响。
结果：发现UCHL3在HCC组织中具有高表达水平。来自60例HCC患者的组织样本用于评估UCHL3和EEF1A1之间的相关性。UCHL3通过赖氨酸位点与EEF1A1结合，这降低了EEF1A1的泛素化水平。功能实验和裸鼠模型已经证明UCHL3/EEF1A1轴促进迁移,stemness,和肝癌细胞的耐药性。降低EEF1A1的表达可以逆转UCHL3对HCC细胞恶性行为的影响。
结论：我们的发现表明UCHL3通过去泛素化结合并稳定EEF1A1。UCHL3和EEF1A1形成促进HCC恶性进展的功能轴，为肝癌的抗肿瘤靶向治疗提供新的见解。
