Warfarin and bevacizumab suppress tumor progression in pancreatic ductal adenocarcinoma by targeting EGFR-PI3K-Akt signaling - inhibition of proliferation/migration and apoptosis induction

Q: What is the main therapeutic rationale for combining warfarin with bevacizumab in PDAC?

Bevacizumab inhibits VEGF-driven angiogenesis, while warfarin has reported anti-tumor activity linked to the Gas6/Axl-PI3K-Akt axis. Because EGFR converges on PI3K-Akt, co-targeting angiogenic and oncogenic signaling was expected to enhance efficacy.

Q: Which methods were used to identify targets and validate effects experimentally?

Network pharmacology and molecular docking were used to identify key targets and predict warfarin-target binding. In vitro assays including CCK-8, wound healing, flow cytometry, qRT-PCR, and Western blot evaluated proliferation, migration, apoptosis, and EGFR-PI3K-Akt pathway gene/protein expression.

Q: What outcomes did the warfarin–bevacizumab combination produce in PDAC cells?

The combination showed optimal anti-proliferative synergy, reduced wound closure compared with blank, and increased apoptosis. It also suppressed EGFR and PI3K at both mRNA and protein levels while supporting apoptosis-related changes including Bcl-2 suppression and Bax/caspase-3 upregulation.

Sep 30, 2026

Uploaded by Stanley

Warfarin combined with bevacizumab was investigated as a synergistic strategy against pancreatic ductal adenocarcinoma, focusing on EGFR-PI3K-Akt signaling suppression. PDAC background highlights aggressive progression, stromal remodeling, chemotherapy resistance, and tumor-associated hypercoagulability, with limited bevacizumab efficacy due to compensatory pathway activation. Using network pharmacology and molecular docking to define overlapping targets, functional assays assessed proliferation, migration, apoptosis, and EGFR-PI3K-Akt pathway expression. The combination reduced growth and migration, enhanced apoptosis, suppressed EGFR/PI3K at mRNA/protein levels, and supported mitochondrial apoptotic activation.

LanguageEnglish

CategoryHealthcare

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Pages16

Reading Time40 min

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Warfarin and bevacizumab suppress tumor progression in pancreatic ductal adenocarcinoma by targeting EGFR-PI3K-Akt signaling: inhibition of proliferation/migration and apoptosis induction Jingjing Chen1\#, Jianjie Ju1\#, Jingting Wang2, Limei Yang2^ 1School of Pharmacy, Fujian Medical University, Fuzhou, China; 2Department of Pharmacy, Provincial Clinical College of Fujian Medical University/Fujian Provincial Hospital/Fuzhou University Affiliated Provincial Hospital, Fuzhou, China Contributions: (I) Conception and design: J Chen, L Yang; (II) Administrative support: L Yang; (III) Provision of study materials or patients: J Wang, L Yang; (IV) Collection and assembly of data: J Chen, J Ju; (V) Data analysis and interpretation: J Chen, J Ju; (VI) Manuscript writing: All authors; (VII) Final approval of manuscript: All authors. \#These authors contributed equally to this work as co-first authors. Correspondence to: Limei Yang, MS. Department of Pharmacy, Provincial Clinical College of Fujian Medical University/Fujian Provincial Hospital/ Fuzhou University Affiliated Provincial Hospital, Fuzhou 350001, China. Email: yanglimei_ [214@163.com](214@163.com). Background: Pancreatic ductal adenocarcinoma (PDAC) exhibits aggressive progression, dense stromal remodeling, and resistance to chemotherapy, resulting in extremely poor survival. Although bevacizumabmediated vascular endothelial growth factor (VEGF) inhibition can suppress angiogenesis, clinical efficacy is limited by compensatory activation of alternative signaling pathways. Meanwhile, PDAC-associated hypercoagulability supports tumor progression, and warfarin has been reported to exert anti-tumor effects partly through inhibition of the growth arrest-specific protein 6 (Gas6)/Axl-phosphoinositide 3-kinase (PI3K)-protein kinase B (Akt) axis. Given that epidermal growth factor receptor (EGFR) also converges on the PI3K-Akt pathway, combining warfarin with bevacizumab may enhance therapeutic efficacy by cotargeting angiogenic and oncogenic signaling. This study aimed to evaluate the synergistic effects of warfarin and bevacizumab in PDAC and to elucidate the underlying molecular mechanisms. Methods: An integrated approach combining network pharmacology, molecular docking, and in vitro assays was used. Bioinformatics tools identified key targets and pathways, with docking simulations assessing warfarin-target binding. Functional assays, including the Cell Counting Kit-8 (CCK-8), wound healing, flow cytometry, quantitative real-time polymerase chain reaction (qRT-PCR), and Western blot, evaluated cell proliferation, migration, apoptosis, and gene/protein expression related to EGFR-PI3K-Akt pathway. Results: Integrated bioinformatics identified 70 overlapping targets between warfarin and pancreatic cancer, with EGFR, PI3K isoforms, and AKT1 as core hubs in the protein-protein interaction (PPI) network. Molecular docking demonstrated strong warfarin binding to EGFR, PI3K catalytic isoforms, and AKT1 (ΔG <−7.0 kcal/mol), while exhibiting moderate interaction with the PI3K regulatory subunit phosphoinositide- 3-kinase regulatory subunit 1 (PIK3R1, −6.1 kcal/mol) . In vitro validation showed that 0.8 mmol/L warfarin combined with 500 mg/L bevacizumab exhibited optimal anti-proliferative synergy (24-h Bliss score: 0.335; 48-h inhibition: 50.5%), while reducing wound closure versus blank (P<0.001) . The combination elevated apoptosis to 10.11%(P<0.001 vs. 1.19% blank) with B-cell lymphoma 2 (Bcl-2) suppression (0.41-fold), Bcl-2-associated X protein (Bax, 1.12-fold) and cysteine-aspartic acid protease-3 (caspase-3, 0.93-fold) upregulation. Combination therapy synergistically downregulated EGFR (0.41-vs. 0.74-fold warfarin, P<0.001) and PI3K (0.32-vs. 0.57-fold, P<0.001) at messenger RNA (mRNA)/protein levels, while ^ ORCID: 0000-0002-6486-9077. © AME Publishing Company. Transl Cancer Res 2025;14(12):8616-8631 | [https://dx.doi.org/10.21037/tcr-2025-1190](https

FAQs

What is the main therapeutic rationale for combining warfarin with bevacizumab in PDAC?

Bevacizumab inhibits VEGF-driven angiogenesis, while warfarin has reported anti-tumor activity linked to the Gas6/Axl-PI3K-Akt axis. Because EGFR converges on PI3K-Akt, co-targeting angiogenic and oncogenic signaling was expected to enhance efficacy.

Which methods were used to identify targets and validate effects experimentally?

Network pharmacology and molecular docking were used to identify key targets and predict warfarin-target binding. In vitro assays including CCK-8, wound healing, flow cytometry, qRT-PCR, and Western blot evaluated proliferation, migration, apoptosis, and EGFR-PI3K-Akt pathway gene/protein expression.

What outcomes did the warfarin–bevacizumab combination produce in PDAC cells?

The combination showed optimal anti-proliferative synergy, reduced wound closure compared with blank, and increased apoptosis. It also suppressed EGFR and PI3K at both mRNA and protein levels while supporting apoptosis-related changes including Bcl-2 suppression and Bax/caspase-3 upregulation.

Sep 30, 2026

Uploaded by Stanley