PRDX1的O-连接N-乙酰葡糖胺化修饰增强其稳定性，并通过激活LRP6介导的Wnt信号通路促进肝癌进展。

O-GlcNAcylation of PRDX1 enhances its stability and promotes liver cancer progression via activating LRP6-mediated Wnt signaling.

作者信息

Dai Luo, Xu Zheng, Fan Yin, Gao Wei, Yuan Guandou, He Songqing, Mao Linfeng

机构信息

Department of Hepatobiliary Surgery, The First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi, China; Key Laboratory of Early Prevention and Treatment for Regional High Frequency Tumor (Guangxi Medical University), Ministry of Education, Nanning 530021, Guangxi, China; Guangxi Key Laboratory of Immunology and Metabolism for Liver Diseases, Nanning 530021, Guangxi, China.

出版信息

Cell Signal. 2025 Aug 7;135:112055. doi: 10.1016/j.cellsig.2025.112055.

DOI:10.1016/j.cellsig.2025.112055

PMID:40783149

Abstract

BACKGROUND

O-GlcNAcylation, a post-translational modification intricately implicated in oncogenic processes, has garnered significant attention as a potential therapeutic target in cancer biology. Peroxiredoxin 1 (PRDX1), a master regulator of reactive oxygen species (ROS) homeostasis and antioxidant defense systems, is increasingly recognized for its contributory role in the pathogenesis of diverse malignancies. However, the functional significance of PRDX1 in liver cancer pathogenesis and the mechanistic underpinnings of its regulation remain to be fully elucidated.

METHODS

In our preliminary investigations, we identified PRDX1 as a substrate amenable to O-GlcNAcylation via immunoprecipitation-mass spectrometry (IP-MS) profiling. Western blotting was performed to determine the levels of PRDX1 and O-GlcNAcylation in liver cancer tissues. Colony formation, scratch test, transwell assay and nude mouse tumor model assays were used to determine the roles of PRDX1 and O-GlcNAcylation in liver cancer progression. IP-MS was used to screen the interacting protein LRP6 of PRDX1, cycloheximide (CHX) chase assay, ubiquitination test were used to determine the stability, proximity ligation assay (PLA), immunofluorescent staining (IF) were performed the O-GlcNAcylation of PRDX1.

RESULTS

Herein, we demonstrate that PRDX1 exerts profound oncogenic effects, driving liver cancer progression in both in vitro and in vivo experimental models. Notably, we reveal that PRDX1 undergoes pronounced O-GlcNAcylation in liver cancer, a modification that enhances its protein stability by attenuating ubiquitin-proteasomal degradation. Furthermore, PRDX1 interacts with low-density lipoprotein receptor-related protein 6 (LRP6), stabilizing its expression and subsequently activating the canonical Wnt/β-catenin signaling cascade.

CONCLUSION

Our findings suggest that O-GlcNAcylation stabilizes PRDX1, promoting liver cancer progression. PRDX1-LRP6 interaction activates Wnt/β-catenin signaling, driving tumorigenesis. Targeting the O-GlcNAcylation-PRDX1-LRP6 axis holds therapeutic promise.

摘要

背景

O-连接的N-乙酰葡糖胺糖基化（O-GlcNAcylation）是一种与致癌过程密切相关的翻译后修饰，作为癌症生物学中的潜在治疗靶点已引起广泛关注。过氧化物酶1（PRDX1）是活性氧（ROS）稳态和抗氧化防御系统的主要调节因子，其在多种恶性肿瘤发病机制中的作用日益受到认可。然而，PRDX1在肝癌发病机制中的功能意义及其调控的分子机制仍有待充分阐明。

方法

在我们的初步研究中，通过免疫沉淀-质谱（IP-MS）分析，我们确定PRDX1是一种适合O-GlcNAcylation修饰的底物。采用蛋白质免疫印迹法检测肝癌组织中PRDX1和O-GlcNAcylation的水平。通过集落形成实验、划痕实验、Transwell实验和裸鼠肿瘤模型实验，确定PRDX1和O-GlcNAcylation在肝癌进展中的作用。利用IP-MS筛选PRDX1的相互作用蛋白低密度脂蛋白受体相关蛋白6（LRP6），采用环己酰亚胺（CHX）追踪实验、泛素化实验确定PRDX1的稳定性，通过邻近连接分析（PLA）、免疫荧光染色（IF）检测PRDX1的O-GlcNAcylation修饰。

结果

在此，我们证明PRDX1在体外和体内实验模型中均发挥着深刻的致癌作用，推动肝癌进展。值得注意的是，我们发现PRDX1在肝癌中发生显著的O-GlcNAcylation修饰，这种修饰通过减弱泛素-蛋白酶体降解来增强其蛋白质稳定性。此外，PRDX1与低密度脂蛋白受体相关蛋白6（LRP6）相互作用，稳定其表达并随后激活经典的Wnt/β-连环蛋白信号级联反应。