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Integrative proteomics and n-glycoproteomics reveal the synergistic anti-tumor effects of aspirin- and gemcitabine-based chemotherapy on pancreatic cancer cells

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Abstract

Objective and design

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.

Methods and results

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.

Conclusions

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.

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Data Availability

The data used and/or analyzed during the present study are available.

Abbreviations

PDAC:

Pancreatic ductal adenocarcinoma

NSAIDs:

Non-steroidal anti-inflammatory drugs

COX-2:

Cyclooxygenase-2

NF-κB:

Nuclear factor kappa-B

TNF:

Tumor necrosis factor

ECM:

Extremely dense extracellular matrix

EMT:

Mesenchymal transition

TME:

Tumor microenvironment

TMT:

Tandem mass tag

KEGG:

Kyoto Encyclopedia of Genes and Genomes

GO:

Gene ontology

INGPs:

Intact N-glycopeptides

IC50:

Half maximal inhibitory concentration

GTs:

Glycosyltransferases

PTM:

Post-translational modification

PPI:

Protein-protein interaction

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Funding

This work was supported by the National Key Program for Basic Research of China (grant numbers 2020YFC2002700, 2020YFE0202200); and Research Program of the State Key Laboratory of Proteomics [grant number SKLP-K201901].

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Author notes

  1. Xiaoyu Li, Ran Kong contributed to the work equally and should be regarded as co-first authors.

Authors and Affiliations

  1. State Key Laboratory of Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Beijing Institute of Lifeomics, No. 38 Life Park Road, Changping District, Beijing, 102206, China

    Xiaoyu Li, Ran Kong, Wenhao Hou, Junxia Cao, Xiaohong Qian & Wantao Ying

  2. Institute of Analysis and Testing, Beijing Center for Physical & Chemical Analysis), Beijing Academy of Science and Technology, Beijing, 100094, China

    Xiaoyu Li

  3. Biomedical Engineering Department, Peking University, Beijing, 100191, China

    Ran Kong

  4. Center for Bioinformatics and Computational Biology, School of Life Sciences, Institute of Biomedical Sciences, East China Normal University, Shanghai, China

    Li Zhang

  5. Beijing Key Laboratory of Environmental and Viral Oncology, Faculty of Environment and Life, Beijing University of Technology, No. 100 Ping Le Yuan, Chaoyang District, Beijing, 100124, China

    Lijiao Zhao

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  1. Xiaoyu Li
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  2. Ran Kong
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  3. Wenhao Hou
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Contributions

X.L. and R.K. contributed to the work equally and should be regarded as co-first authors.X.L : Conceptualization; Experimenter; Data curation; Formal analysis; Visualization; Roles/Writing original draft;R.K : Investigation; Roles/Writing original draft; Experimenter;W.H : Writing editing;L.Z : Formal analysis; J.C : Experimenter;X.Q : Project administration; Resources; Funding acquisition;L.Z : Project administrationW.Y : Project administration; Resources; Funding acquisition; Writing review & editing.

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Correspondence to Lijiao Zhao or Wantao Ying.

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Li, X., Kong, R., Hou, W. et al. Integrative proteomics and n-glycoproteomics reveal the synergistic anti-tumor effects of aspirin- and gemcitabine-based chemotherapy on pancreatic cancer cells. Cell Oncol. 47, 141–156 (2024). https://doi.org/10.1007/s13402-023-00856-z

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