Modulating Wnt/β-Catenin Signaling Pathway on U251 and T98G Glioblastoma Cell Lines Using a Combination of Paclitaxel and Temozolomide, A Molecular Docking Simulations and Gene Expression Study

Abstract

One of the most lethal cancers, glioblastoma (GBM), affects 14.5 of all central nervous system (CNS) tumors. Patients diagnosed with GBM have a meager median overall survival (OS) of 15 months. Extensive genetic analysis has shown that many dysregulated pathways, including the Wnt/beta-catenin signaling system, contribute to the pathogenicity of GBM. Paclitaxel (PTX) and temozolomide (TMZ) are recognized to have therapeutic potential in several types of cancer, including GBM. This work aimed to examine the impact of PTX and TMZ on the human glioma cell lines U251 and T98G using molecular docking simulations and gene expression profiles in the Wnt/beta-catenin signaling pathway. Standard procedure for Molecular Docking simulation, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity assay, and Flow Cytometry assay was used. Genes implicated in the Wnt/beta-catenin signaling pathway, including Dvl, Axin, APC, beta-catenin, and glycogen synthase kinase3-beta (GSK3 beta), were subjected to real-time PCR. The estimated parameters for targets revealed that the average binding energy and inhibition constant (Ki) for the DVL, beta-Catenin, and GSK3 beta, when targeted by PTX, were -5.01 kcal/mol, -5.4 kcal/mol, and -9.06 kcal/mol, respectively. This energy range was -6.34 kcal/mol for DVL, -5.52 kcal/mol for beta-Catenin, and -5.66 kcal/mol for GSK3 beta as a result of TMZ's inhibitory actions. Gene expression analyses indicated that PTX and PTX/TMZ suppressed GSK3 beta (p < 0.05). GSK3fl from the Wnt/beta-catenin signaling pathway was significantly targeted by PTX alone, and adding TMZ to PTX may improve the efficacy of glioblastoma treatment. In addition, the GSK3 beta gene may help GBM therapy strategies as a potential PTX target.