化疗耐药性很大程度上阻碍了化疗药物在癌症患者中的临床使用，联合或序贯药物治疗方案的设计目的是尽量减少化疗毒性并使化疗耐药性重新敏感。在这项工作中，发现 HeLa 细胞中帕博西尼预处理可增强顺铂的细胞毒性作用。通过基于液相色谱-质谱法的蛋白质组学和N-糖蛋白质组学工作流程的整合，我们发现单独的palbociclib主要增强HeLa细胞中的N-糖基化改变，而顺铂主要增加与凋亡途径相关的不同表达蛋白。结果，两种药物的连续使用诱导了凋亡蛋白BAX和BAK的较高表达水平。 palbociclib 诱导的那些改变的 N-糖蛋白涉及与细胞膜修饰和药物敏感性密切相关的途径。具体而言，前四种常见糖基化蛋白 FOLR1、L1CAM、CD63 和 LAMP1 均与耐药性或药物敏感性相关。据怀疑，palbociclib 诱导的膜蛋白 N-糖基化使 HeLa 细胞变得更容易受到顺铂治疗。我们的研究为靶向药物和化疗药物顺序使用的机制提供了新的见解，同时提出了一种涉及蛋白质组学和 N-糖蛋白质组学的高效方法，以促进药物发现。版权所有 © 2024 作者。由 Wolters Kluwer Health, Inc. 出版

Chemoresistance largely hampers the clinical use of chemodrugs for cancer patients, combination or sequential drug treatment regimens have been designed to minimize chemotoxicity and resensitize chemoresistance. In this work, the cytotoxic effect of cisplatin was found to be enhanced by palbociclib pretreatment in HeLa cells. With the integration of liquid chromatography-mass spectrometry-based proteomic and N-glycoproteomic workflow, we found that palbociclib alone mainly enhanced the N-glycosylation alterations in HeLa cells, while cisplatin majorly increased the different expression proteins related to apoptosis pathways. As a result, the sequential use of two drugs induced a higher expression level of apoptosis proteins BAX and BAK. Those altered N-glycoproteins induced by palbociclib were implicated in pathways that were closely associated with cell membrane modification and drug sensitivity. Specifically, the top four frequently glycosylated proteins FOLR1, L1CAM, CD63, and LAMP1 were all associated with drug resistance or drug sensitivity. It is suspected that palbociclib-induced N-glycosylation on the membrane protein allowed the HeLa cell to become more vulnerable to cisplatin treatment. Our study provides new insights into the mechanisms underlying the sequential use of target drugs and chemotherapy drugs, meanwhile suggesting a high-efficiency approach that involves proteomic and N-glycoproteomic to facilitate drug discovery.Copyright © 2024 The Author(s). Published by Wolters Kluwer Health, Inc.