ORIGINAL RESEARCH article
Front. Immunol.
Sec. Cancer Immunity and Immunotherapy
Volume 16 - 2025 | doi: 10.3389/fimmu.2025.1664468
This article is part of the Research TopicMicrobiome-Driven Modulation of Cancer Therapies: Insights into Chemotherapy, Immunotherapy, and Metabolomic ApproachesView all articles
Combination Therapy with Nisin, Urolithin B, and Vincristine Exhibits Synergistic Antiproliferative and Pro-Apoptotic Effects Against Human Lymphoma Cells: Evidence From Proteomics
Provisionally accepted
- 1NICM Health Research Institute, Western Sydney University, Westmead, Australia
- 2Cairo University Faculty of Pharmacy, Cairo, Egypt
- 3Pharmacology Unit, School of Medicine, Western Sydney University, Campbelltown, Australia
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Lymphoma remains a significant global health burden, necessitating innovative, targeted therapeutic strategies. Nisin (N), a bacteriocin produced by Lactococcus lactis, has demonstrated antimicrobial and anticancer properties through membrane disruption and apoptotic induction. Urolithin B (UB), a gut microbiota-derived metabolite of ellagitannins, has shown anti-inflammatory and antiproliferative activities in various cancer models. Vincristine (Vinc), a common anti-lymphoma drug, disrupts microtubule formation, leading to cell cycle arrest and apoptosis in cancer cells. This study explored the antiproliferative and pro-apoptotic effects of a triple combination therapy comprising N, UB, and Vinc against human lymphoma cell lines (HKB-11 and Hs 313.T). This study systematically evaluated the synergistic efficacy of both monotherapy and dual and triple combinations and molecular mechanisms using Alamar Blue viability assays, combination index (CI) modelling, reactive oxygen species (ROS) quantification, annexin V/7-AAD flow cytometry, and bottom-up label-free proteomics. The potential cytotoxicity of the combination on normal stromal HS-5 cells was also assessed using the Alamar Blue assay. The N: UB: Vinc combination at 2240: 210: 0.94 µM demonstrated potent synergy (CI values 0.31–0.50 at IC90 - IC95) and induced near-complete growth inhibition (> 99%) in both lymphoma cell lines with reduced toxicity (42.09 ± 1.21% viability) toward normal stromal HS-5 cells. ROS analysis revealed significant oxidative stress, while flow cytometry confirmed enhanced apoptosis (p < 0.0001) in the combination groups. Proteomic profiling of the combination N: UB: Vinc at 2450.94 µM uncovered distinct molecular responses, including upregulation of MAP1LC3B2 (Log2FC = 1.4), GMNN (Log2FC = 1.3), and SLC38A2 (Log2FC = 1.5), promoting apoptosis, cell cycle regulation, and mTOR signalling inhibition. Concurrently, key oncogenic and metabolic proteins were downregulated, including NNMT (Log2FC = –2.9), PLTP (Log2FC = –2.5), and CYP4X1 (Log2FC = –2.0), which implicated the suppression of MAPK-Akt signalling, ferroptosis activation, and lipid metabolism disruption. These results established a mechanistic rationale for combining postbiotics such as N and UB with standard chemotherapy, highlighting a promising avenue for safer and more effective lymphoma management in the future.
Keywords: Lymphoma, Vincristine, Nisin, Urolithin B, Synergy, Proteomics, Apoptosis
Received: 12 Jul 2025; Accepted: 04 Sep 2025.
Copyright: © 2025 Al-Khazaleh, Alsherbiny, Chang, Muench and Bhuyan. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Ahmad Al-Khazaleh, NICM Health Research Institute, Western Sydney University, Westmead, Australia
Deep Jyoti Bhuyan, NICM Health Research Institute, Western Sydney University, Westmead, Australia
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