The effect of combination therapy on Bevacizumab, Carmustine, and Metformin chemotherapy drugs on the fluctuating performance expression of TLR2, TLR6, and IL-6 genes in the cell line of brain glioblastoma.

OBJECTIVE: Glioblastoma, one of the most aggressive brain tumors, is distinguished by its resistance to standard treatments. So, studying practical therapeutic methods is of great importance. In this project, the effects of combined chemotherapy drugs, including bevacizumab, carmustine, and metformin, on the expression of IL-6, TLR2, and TLR6 in the glioblastoma cell line of U87MG were investigated. METHODS: U87MG cells were treated with bevacizumab, carmustine, and metformin alone or in combined groups. Cell viability was measured using the MTT assay. Apoptotic rates and cell cycle were analyzed by flow cytometry. Gene and protein expression levels of TLR2, TLR6, and IL-6 were evaluated by Real-Time PCR and Western blotting, respectively. In addition, bioinformatic analyses, including protein-protein interaction (PPI) network construction and pathway enrichment, were performed using Cytoscape (GeneMANIA) and R (ClusterProfiler). RESULTS: The triple-drug combination(bevacizumab, carmustine, and metformin )significantly reduced cell viability and induced a substantial increase in apoptosis and G0/G1 cell cycle arrest. Treatment also led to a marked downregulation of TLR2, TLR6, and IL-6 gene expression. Bioinformatic analysis revealed these genes to be central in pathways associated with immune regulation, inflammation, and tumor progression. CONCLUSION: The combination of bevacizumab, carmustine, and metformin exerts a potent synergistic antitumor effect in U87MG glioblastoma cells through modulation of inflammation-associated genes, induction of apoptosis, and disruption of cell cycle progression. These findings provide a mechanistic rationale for multi-targeted combination therapy in glioblastoma and support further preclinical evaluation.