高等学校化学学报

高等学校化学学报

• 研究论文 • 上一篇    

化学酶法解析乳腺癌细胞治疗性早衰进程中线粒体蛋白质O-GlcNAc修饰时序性改变

蒋馨雅1#，马起乐2#，程洪英3，刘  寅3，黄  煌2,4，赵  然2，刘宇博2，仲小敏1,3   

  1. 1. 南京医科大学附属淮安第一医院肿瘤科

    2. 大连理工大学，生命科学与药学系

    3. 徐州医科大学附属淮安临床学院肿瘤科 4. 艾美诚信生物制药有限公司博士后工作站

  • 收稿日期:2025-04-23 修回日期:2025-05-22 网络首发:2025-05-23 发布日期:2025-05-23
  • 通讯作者: 刘宇博 E-mail:liuyubo@dlut.edu.cn
  • 基金资助:
    徐州医科大学附属医院科技发展基金(批准号：XYFZ202204)和江苏省研究生实践创新计划(批准号：SJCX24_1559)资助

Chemical Enzymatic Analysis of Temporal Alterations in Mitochondrial Protein O - GlcNAc Modification during the Therapeutic Senescence Process of Breast Cancer Cells

JIANG Xinya1#, MA Qile2#, CHENG Hongying3, LIU Yin3, HUANG Huang, ZHAO Ran, LIU Yubo2, ZHONG Xiaomin1,3   

  1. 1. Department of Oncology, The First Affiliated Hospital of Huai’an, Nanjing Medical University

    2. Dalian University of Technology, Department of Life Sciences and Pharmacy

    3. Department of Oncology, Huai’an Clinical College Affiliated to Xuzhou Medical University 4. Postdoctoral workstation of Dalian Hissen Bio Pharm, Inc

  • Received:2025-04-23 Revised:2025-05-22 Online First:2025-05-23 Published:2025-05-23
  • Contact: Yubo Liu E-mail:liuyubo@dlut.edu.cn
  • Supported by:

    Supported by the Science and Technology Development Fund of the Affiliated Hospital of Xuzhou Medical University (No.XYFZ202204) and the Jiangsu Postgraduate Practice Innovation Program (No.SJCX24_1559)

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摘要： 细胞早性衰老与线粒体功能障碍密切相关. 本研究以阿霉素诱导乳腺癌细胞MCF-7构建治疗性早性衰老细胞模型，利用基于GalT1酶（Y289L）的化学酶法标记早衰进程中多个时间点的MCF-7细胞线粒体O-GlcNAc糖蛋白质组，利用点击化学反应正交捕获线粒体糖蛋白，通过LC-MS/MS进行非标定量蛋白质组学分析，研究细胞早性衰老进程中线粒体O-GlcNAc糖蛋白的时序性定量变化，并对其进行生物学功能富集解析，寻找参与调控乳腺癌细胞早性衰老的线粒体O-GlcNAc修饰蛋白，解析线粒体O-GlcNAc糖基化调控细胞早性衰老的机制.

关键词: 化学酶标记, 细胞早性衰老, O-GlcNAc修饰, 蛋白质组学

Abstract: Cellular premature senescence is closely related to mitochondrial dysfunction. In this study, doxorubicin was used to induce breast cancer cells MCF-7 to establish a model of therapeutic premature senescence cells. The chemical enzymatic method based on GalT1 enzyme (Y289L) was employed to label the mitochondrial O-GlcNAc glycoproteome of MCF-7 cells at multiple time points during the premature senescence process. Click chemistry reaction was utilized to orthogonally capture mitochondrial glycoproteins. Label-free quantitative proteomics analysis was carried out through LC-MS/MS to investigate the temporal quantitative changes of mitochondrial O-GlcNAc glycoproteins during the cellular premature senescence process. Moreover, the biological function enrichment analysis was conducted to identify mitochondrial O-GlcNAc modified proteins involved in the regulation of premature senescence of breast cancer cells, and to elucidate the mechanism by which mitochondrial O-GlcNAc glycosylation regulates cellular premature senescence.

Key words: Chemoenzymatic labelling methods, Cell senescence, O-GlcNAcylation, Proteomics

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