Open Access

ARTICLE

Epibrassinolide Induces Apoptosis and Inhibits the Migration of Gastric Cancer AGS Cells by Regulating Reactive Oxygen Species-Mediated Signaling Pathways

Chang Wang^1,2,#, Zhi Zhang^1,#, Wei Sun¹, Quan Quan³, Wenshuang Hou³, Chenghao Jin^1,3,4,*

1 Department of Food Science and Engineering, College of Food Science, Heilongjiang Bayi Agricultural University, Daqing, 163319, China
2 Agricultural Products and Processed Products Supervision and Testing Center, Ministry of Agriculture, Daqing, 163319, China
3 Department of Biochemistry and Molecular Biology, College of Life Science and Technology, Heilongjiang Bayi Agricultural University, Daqing, 163319, China
4 National Coarse Cereals Engineering Research Center, Daqing, 163319, China

* Corresponding Author: Chenghao Jin. Email:
# These authors contributed equally to this work

BIOCELL 2025, 49(3), 465-482. https://doi.org/10.32604/biocell.2025.062155

Received 11 December 2024; Accepted 05 March 2025; Issue published 31 March 2025

Abstract

Objectives: Epibrassinolide (EBR) is a steroid hormone with anti-tumor properties. Nevertheless, its potential to inhibit gastric cancer (GC) cells remains unknown. The aim of this research was to examine the effects of EBR on GC cells and to investigate the specific mechanism of EBR. Methods: A cell counting kit-8 (CCK-8) assay was utilized to determine cell survival rates. The investigation of apoptosis, cell cycle progression, and reactive oxygen species (ROS) levels was performed using flow cytometry. To detect cell migration, a wound-healing assay was performed on AGS cells. Furthermore, western blotting assay was utilized to determine protein expression levels. Results: The CCK-8 assay demonstrated that EBR reduced the survival rates of AGS, KATO-3, and MKN-45 cells, while causing only minor toxicity to normal cells. The apoptosis assay indicated that EBR induced AGS cell apoptosis through a mitochondria-mediated pathway. Western blotting results demonstrated that EBR induced AGS cell apoptosis via mitogen-activated protein kinase (MAPK)/signal transducer and activator of transcription 3 (STAT3)/nuclear factor kappa B (NF-κB) signaling pathway. Further, after treating AGS cells with EBR, the accumulation of intracellular ROS markedly increased. EBR also induced G2/M phase cell cycle arrest in AGS cells by downregulating phospho-protein kinase B (p-AKT), cyclin-dependent kinase 1/2 (CDK1/2), and cyclin B1 expression levels, while simultaneously upregulating p21 and p27 expression levels. EBR inhibited AGS cell migration by downregulating p-AKT, phosphorylated-glycogen synthase kinase 3β (p-GSK-3β), and β-catenin expression levels and upregulating E-cadherin expression levels. However, these effects were reversed by pretreatment with N-acetylcysteine (NAC). Conclusion: EBR regulates AGS cells by inducing apoptosis and G2/M phase arrest, while also inhibiting cell migration, all of which are mediated through ROS-mediated signaling pathways. Ultimately, these effects suggest a significant role for EBR in regulating cellular processes within AGS cells.

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