Diffuse Large B‑Cell Lymphoma (DLBCL) is the most common and aggressive subtype of non‑Hodgkin lymphoma, notable for clinical heterogeneity and frequent chemoresistance. Approximately 30% of patients experience relapse or develop refractory disease despite standard therapies. To address this unmet need, the authors applied a drug repurposing strategy that integrates computational network pharmacology, molecular docking, and in vitro experimentation with the aim of identifying existing antitumor agents with potential synergistic efficacy against DLBCL.
DLBCL‑associated genes were retrieved from DisGeNET and GeneCards. Overlapping genes (OGs) between sources were identified using Venny v2.1.0, yielding 144 OGs that formed the basis for downstream analyses. Functional enrichment of these OGs was performed with ShinyGO v0.77 to determine implicated biological processes and pathways.
Protein‑protein interaction (PPI) networks were generated using STRING v11.5 and visualized in Cytoscape v3.10 to identify hub genes within the DLBCL‑associated network. Expression validation of selected hubs was carried out using GEPIA2.
Potential therapeutic compounds were screened from DSigDB and DrugMAP databases. Molecular docking of candidate drugs to hub gene proteins was performed using AutoDock Vina v1.2.0 to estimate binding affinities and prioritize repurposing candidates for experimental testing.
In vitro validation used the 2PK‑3 DLBCL cell line to assess cytotoxicity and oxidative stress. Cells were treated with Doxorubicin and Paclitaxel, both as single agents and in combination. Morphological assessment, MTT viability assays, and reactive oxygen species (ROS) measurements were used to evaluate treatment effects.
Docking analysis identified several compounds with high predicted binding affinity (reported as ≤ −8.0 kcal/mol in the abstract). Among these, Doxorubicin, Paclitaxel, Sorafenib, Masoprocol, and Bortezomib demonstrated favorable docking scores against the selected hub proteins. The study highlights that these docking results supported selection of Doxorubicin and Paclitaxel for in vitro evaluation.
Note: the abstract reports affinity thresholds and lists top compounds; the full text is required for detailed per‑target docking scores and binding site interactions, which are not provided in the abstract.
The authors tested Doxorubicin and Paclitaxel on cultured 2PK‑3 DLBCL cells to compare single‑agent and combined effects. Endpoints included cell viability measured by MTT assay, microscopic morphological analysis to detect cellular disruption, and assays quantifying intracellular ROS as an indicator of oxidative stress.
Combined treatment resulted in enhanced cytotoxicity relative to either drug alone. Morphological disruption was more pronounced with the combination, and ROS generation was elevated compared with single‑agent treatments. These in vitro observations are presented as supporting evidence for potential synergy between Doxorubicin and Paclitaxel in this DLBCL model.
The integrative workflow—from curated disease genes through enrichment, network analysis, docking, and cell‑based validation—supports the identification of repurposed drugs with potential combinatorial efficacy. The study presents evidence that combining two established antitumor agents, Doxorubicin and Paclitaxel, may exert synergistic effects in DLBCL models by engaging pathways such as PI3K‑Akt and apoptosis and by increasing oxidative stress in tumor cells.
These findings suggest that systematic repurposing approaches can help prioritize existing drugs for combination regimens in DLBCL, potentially accelerating translational development compared with de novo drug discovery.
The abstract summarizes methods and principal results but omits several specifics required for full appraisal and reproducibility: exact numerical docking scores per target, concentrations and exposure durations used in cell culture assays, statistical analyses, effect sizes, and any toxicity assessments. The abstract also does not report whether combination effects were formally tested for synergy using methods such as combination index calculations. For these details, consultation of the full text is necessary.
This study reports an integrative drug repurposing pipeline that identified Doxorubicin and Paclitaxel as candidate agents with enhanced combined activity against DLBCL in vitro. The results support further investigation of this combination and illustrate how computational and experimental methods can be combined to prioritize repurposed therapeutics for aggressive lymphomas. The authors declared no competing interests. For full methodological detail and quantitative results, readers should refer to the full published article.