Vincristine exerts antiglioma effects by inhibiting the PI3K/AKT signaling pathway: A mechanistic study based on network pharmacology, bioinformatics analysis, and experimental validation

Abstract

In clinical settings, glioma patients often develop secondary resistance to first-line chemotherapy drugs. Vincristine has been reported for its application in cancer chemotherapy, but its molecular mechanism of action remains unclear. This study aimed to identify potential targets of vincristine in glioma using network pharmacology and to experimentally validate the possible molecular mechanisms against glioma. First, the potential targets of vincristine were predicted using CTD, SwissTargetPrediction, and TargetNet databases. Differential expression analysis and WGCNA algorithm were employed on glioma data from the GEO database to obtain important glioma-related target genes, which were then used to identify the anti-glioma targets of vincristine. The intersecting targets were input into the String database to construct a PPI network, and core targets were identified using the cytohubba plugin in Cytoscape. GO and KEGG analyses were conducted to investigate the functional and pathway enrichment of the intersecting targets. The expression and prognostic significance of the core targets were validated using data from the TCGA and HPA databases. Finally, the anti-glioma proliferation effect of vincristine was validated through CCK-8 assay, flow cytometry for cell cycle analysis, RT-qPCR, and Western blotting. A total of 175 vincristine targets and 1673 glioma targets were identified, with 11 shared targets between vincristine and glioma tissues. Network pharmacology studies suggested that CDC25B, CDK4, CDK6, TOP2A, and the PI3K/AKT signaling pathway might be important core targets and pathways through which vincristine exerts its anti-glioma effects. In vitro experiments confirmed that vincristine successfully inhibited U87 cell proliferation and induced G1 phase arrest via the PI3K/AKT signaling pathway, thereby reducing cell growth. The study results indicate that the PI3K/AKT signaling pathway may be involved in the mechanism by which vincristine inhibits the proliferation of glioma cells.

Keywords: Bioinformatics; Glioma; Network Pharmacology; PI3K/AKT Signaling Pathway; Vincristine.