Unraveling the Complexity of Glioblastoma Microenvironment: A Comparative Study of 3D and 2D Cultures in Response to Combination Drug Therapy Publisher Pubmed



R Rashidi RASOUL ; Mr Hadjighasem Mahmoudreza REZA ; B Negahdari BABAK
Source: Brain Research Published:2025

Abstract

Glioblastoma multiforme (GBM) represents one of the most prevalent and treatment-resistant forms of brain cancer. This investigation explored the therapeutic potential of a combined Erlotinib and Imatinib regimen through experimental studies conducted in both two-dimensional and three-dimensional cell culture systems. Cellular viability and apoptotic responses were evaluated 24- and 48-hours post-treatment utilizing MTT assays and flow cytometry, while gene expression analysis of Bcl-2 and VEGF was performed using qRT-PCR. Furthermore, the scratch assay was employed to assess the impact of drug treatment on cellular migration. Notably, lower drug concentrations were required in 3D cultures due to enhanced cell–cell interactions, resulting in greater cytotoxicity and a marked inhibition of tumor cell proliferation. Significant downregulation of Bcl-2 and VEGF expression was observed, particularly a pronounced reduction in Bcl-2, which correlated with elevated apoptosis rates after 48 h. Furthermore, the apoptotic effect of combination therapy was confirmed, with an increase in death percentage in 48 h-3D treated groups, as expected (*P < 0.05 for monotherapy and ***P < 0.001 for combination). The 3D culture model provided a physiologically relevant context for evaluating oncolytic therapies. These findings support the need for continued investigation through advanced preclinical models and eventual clinical validation. © 2025 Elsevier B.V., All rights reserved.