O-GlcNAcylation promotes malignancy and cisplatin resistance of lung cancer by stabilising NRF2

IF 7.9 1区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Clinical and Translational Medicine Pub Date : 2024-10-02 DOI:10.1002/ctm2.70037

Yihan Zhang, Changning Sun, Leina Ma, Guokai Xiao, Yuchao Gu, Wengong Yu

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Abstract

Background

The transcription factor NRF2 plays a significant role in regulating genes that protect cells from oxidative damage. O-GlcNAc modification, a type of posttranslational modification, is crucial for cellular response to stress. Although the involvement of both NRF2 and O-GlcNAc in maintaining cellular redox balance and promoting cancer malignancy has been demonstrated, the potential mechanisms remain elusive.

Methods

The immunoblotting, luciferase reporter, ROS assay, co-immunoprecipitation, and immunofluorescence was used to detect the effects of global cellular O-GlcNAcylation on NRF2. Mass spectrometry was utilised to map the O-GlcNAcylation sites on NRF2, which was validated by site-specific mutagenesis and O-GlcNAc enzymatic labelling. Human lung cancer samples were employed to verify the association between O-GlcNAc and NRF2. Subsequently, the impact of NRF2 O-GlcNAcylation in lung cancer malignancy and cisplatin resistance were evaluated in vitro and in vivo.

Results

NRF2 is O-GlcNAcylated at Ser103 residue, which hinders its binding to KEAP1 and thus enhances its stability, nuclear localisation, and transcription activity. Oxidative stress and cisplatin can elevate the phosphorylation of OGT at Thr444 through the activation of AMPK kinase, leading to enhanced binding of OGT to NRF2 and subsequent elevation of NRF2 O-GlcNAcylation. Both in cellular and xenograft mouse models, O-GlcNAcylation of NRF2 at Ser103 promotes the malignancy of lung cancer. In human lung cancer tissue samples, there was a significant increase in global O-GlcNAcylation, and elevated levels of NRF2 and its O-GlcNAcylation compared to paired adjacent normal tissues. Chemotherapy promotes NRF2 O-GlcNAcylation, which in turn decreases cellular ROS levels and drives lung cancer cell survival.

Conclusion

Our findings indicate that OGT O-GlcNAcylates NRF2 at Ser103, and this modification plays a role in cellular antioxidant, lung cancer malignancy, and cisplatin resistance.

Abstract Image

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O-GlcNAcylation 通过稳定 NRF2 促进肺癌的恶性程度和顺铂耐药性。

背景转录因子 NRF2 在调节保护细胞免受氧化损伤的基因方面发挥着重要作用。O-GlcNAc修饰是一种翻译后修饰，对细胞应激反应至关重要。虽然 NRF2 和 O-GlcNAc 参与维持细胞氧化还原平衡和促进癌症恶性发展的作用已得到证实，但其潜在机制仍难以捉摸：方法：采用免疫印迹、荧光素酶报告、ROS 检测、共免疫沉淀和免疫荧光等方法检测细胞中 O-GlcNAcylation 对 NRF2 的影响。利用质谱法绘制了 NRF2 上的 O-GlcNAcylation 位点图，并通过位点特异性突变和 O-GlcNAc 酶标记进行了验证。人类肺癌样本被用来验证 O-GlcNAc 与 NRF2 之间的关联。随后，在体外和体内评估了 NRF2 O-GlcNAcylation 对肺癌恶性程度和顺铂耐药性的影响：结果：NRF2在Ser103残基上被O-GlcNAcylated，这阻碍了其与KEAP1的结合，从而增强了其稳定性、核定位和转录活性。氧化应激和顺铂可通过激活 AMPK 激酶提高 OGT 在 Thr444 处的磷酸化，从而增强 OGT 与 NRF2 的结合，进而提高 NRF2 O-GlcNAcylation 的水平。在细胞和异种移植小鼠模型中，NRF2 在 Ser103 处的 O-GlcNAcylation 都会促进肺癌的恶性发展。在人类肺癌组织样本中，全局 O-GlcNAcylation 显著增加，与配对的邻近正常组织相比，NRF2 及其 O-GlcNAcylation 水平升高。化疗促进了 NRF2 O-GlcNAcylation 的形成，进而降低了细胞 ROS 水平，推动了肺癌细胞的存活：我们的研究结果表明，OGT O-GlcNA酰化NRF2的Ser103位点，这种修饰在细胞抗氧化、肺癌恶性程度和顺铂抗性中发挥作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Clinical and Translational Medicine Multiple-

CiteScore

15.90

自引率

1.90%

发文量

450

审稿时长

4 weeks

期刊介绍： Clinical and Translational Medicine (CTM) is an international, peer-reviewed, open-access journal dedicated to accelerating the translation of preclinical research into clinical applications and fostering communication between basic and clinical scientists. It highlights the clinical potential and application of various fields including biotechnologies, biomaterials, bioengineering, biomarkers, molecular medicine, omics science, bioinformatics, immunology, molecular imaging, drug discovery, regulation, and health policy. With a focus on the bench-to-bedside approach, CTM prioritizes studies and clinical observations that generate hypotheses relevant to patients and diseases, guiding investigations in cellular and molecular medicine. The journal encourages submissions from clinicians, researchers, policymakers, and industry professionals.