胰腺癌是一种恶性程度高、预后差的肿瘤。我们基于糖基化，进行了一项综合定量N-糖基蛋白质组学研究，以探究阿司匹林和吉西他滨对胰腺癌细胞协同抗肿瘤作用，并探索胰腺癌化疗的潜在分子机制。我们分别使用吉西他滨、阿司匹林和二者的联合（吉西他滨+阿司匹林）来处理两种胰腺癌细胞系（PANC-1和BxPC-3）。我们发现加入阿司匹林增强了吉西他滨对PANC-1和BxPC-3细胞活性的抑制作用。我们通过综合多组学分析获得了定量N-糖基蛋白质组、蛋白质组、磷酸化和转录组数据，评估了阿司匹林和吉西他滨对胰腺癌细胞的抗肿瘤作用。整体N-糖肽质谱分析的Mfuzz结果显示了两个与阿司匹林相关的一致趋势，表明N-糖基化与阿司匹林的协同作用之间存在强烈关系。进一步分析显示，对胰腺癌细胞的胶原基质相关蛋白（LAMP1、LAMP2、ITGA3等）的唾液酸化和高甘露糖形式的动态调控是阿司匹林促进吉西他滨的抗肿瘤活性以及胰腺癌细胞的药物耐药性的重要因素。对胰腺癌细胞中与N-糖基化相关的过程和途径进行深入分析，可以为未来的胰腺癌治疗靶点和药物耐药机制的研究提供新的见解。© 2023. Springer Nature Switzerland AG.

Pancreatic cancer is a highly malignant tumor that is well known for its poor prognosis. Based on glycosylation, we performed integrated quantitative N-glycoproteomics to investigate the synergistic anti-tumor effects of aspirin and gemcitabine on pancreatic cancer cells and explore the potential molecular mechanisms of chemotherapy in pancreatic cancer.Two pancreatic cancer cell lines (PANC-1 and BxPC-3) were treated with gemcitabine, aspirin, and a combination (gemcitabine + aspirin). We found that the addition of aspirin enhanced the inhibitory effect of gemcitabine on the activity of PANC-1 and BxPC-3 cells. Quantitative N-glycoproteome, proteome, phosphorylation, and transcriptome data were obtained from integrated multi-omics analysis to evaluate the anti-tumor effects of aspirin and gemcitabine on pancreatic cancer cells. Mfuzz analysis of intact N-glycopeptide profiles revealed two consistent trends associated with the addition of aspirin, which showed a strong relationship between N-glycosylation and the synergistic effect of aspirin. Further analysis demonstrated that the dynamic regulation of sialylation and high-mannose glycoforms on ECM-related proteins (LAMP1, LAMP2, ITGA3, etc.) was a significant factor for the ability of aspirin to promote the anti-tumor activity of gemcitabine and the drug resistance of pancreatic cancer cells.In-depth analysis of N-glycosylation-related processes and pathways in pancreatic cancer cells can provide new insight for future studies regarding pancreatic cancer therapeutic targets and drug resistance mechanisms.© 2023. Springer Nature Switzerland AG.