Unraveling the complexity of glioblastoma microenvironment: A comparative study of 3D and 2D cultures in response to combination drug therapy.

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.