Open Access

ARTICLE

VCA Augments Doxorubicin Efficacy in Triple-Negative Breast Cancer: Evidence for Multi-Pathway Synergism

Chang-Eui Hong^1,2,3, Su-Yun Lyu^1,2,3,*

1 Department of Pharmacy, Sunchon National University, Suncheon, 57922, Republic of Korea
2 Smart Beautytech Research Institute, Sunchon National University, Suncheon, 57922, Republic of Korea
3 Research Institute of Life and Pharmaceutical Sciences, Sunchon National University, Suncheon, 57922, Republic of Korea

* Corresponding Author: Su-Yun Lyu. Email:

(This article belongs to the Special Issue: Natural Product-Based Anticancer Drug Discovery)

BIOCELL 2025, 49(12), 2377-2397. https://doi.org/10.32604/biocell.2025.072360

Received 25 August 2025; Accepted 30 October 2025; Issue published 24 December 2025

Abstract

Objective: Triple-negative breast cancer (TNBC) remains a major therapeutic challenge with limited treatment options and poor prognosis. This study aimed to investigate the synergistic anticancer effects of doxorubicin (DOX) combined with Viscum album L. var. coloratum agglutinin (VCA) and to elucidate the underlying molecular mechanisms in TNBC cells. Methods: This study evaluated the synergistic effects and mechanisms of doxorubicin (DOX) and Viscum album L. var. coloratum agglutinin (VCA) combination in MDA-MB231 TNBC cells. Cell viability, oxidative stress markers, apoptosis-related proteins, cell migration, and proliferative recovery were assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, superoxide dismutase (SOD) and nitric oxide (NO) assays, Western blotting, wound healing assay, and Muse™ cell analyzer, respectively. Results: The DOX-VCA combination demonstrated strong synergistic cytotoxicity with Bliss Independence scores of +8.9% to +33.4% at therapeutic concentrations (0.01–50 ng/mL, p = 0.032) and remarkable dose reduction indices of >3000-fold for DOX and >16.7-fold for VCA. This synergistic effect was mediated through multiple mechanisms: enhanced oxidative stress modulation (48% increase in SOD-like activity, p = 0.0003, and 94% increase in NO production, p = 0.0002, at 50 ng/mL combination compared to control), augmented apoptotic responses (4.8-fold increase in cleaved caspase-3/caspase-3 ratio, p = 0.0001, and 91% reduction in procaspase-9 levels, p = 0.00008, at 48 h compared to control), significant inhibition of cell migration (85.8% remaining wound area at 48 h, p = 0.0004 vs.control), and severely impaired proliferative recovery (98.9% reduction in cell viability at 72 h post-treatment, p = 0.0001 vs. untreated control). Conclusion: The DOX-VCA combination demonstrates potent synergistic effects through multiple mechanisms, warranting further investigation as a potential dose-reducing strategy for TNBC treatment.

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