The inhibitory effects of Cucurbitacin B on hypopharyngeal cancer cell malignant behaviors were investigated based on network pharmacology and molecular docking

Hypopharyngeal cancer is a metastatic squamous cell carcinoma of the head and neck (SCCHN), and the early symptoms are not obvious. Cucurbitacin B (Cu-B) is the most abundant member of the Cucurbitacin family and has a wide range of pharmacological activities, but its effect on hypopharyngeal cancer is not very clear. The cell functions were detected by the cell counting kit-8 (CCK-8), colony formation assay, 5-ethynyl-2′-deoxyuridine (EdU) staining, flow cytometry, wound healing, transwell, and tube formation assays. The Gene Expression Omnibus (GEO), Comparative Toxicogenomics Database (CTD), SwissTargetPrediction, and GeneCards databases were applied to identify the targets of Cu-B in hypopharyngeal cancer. What’s more, the protein-protein interaction (PPI) network was constructed using the String database, and the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed employing the DAVID database. The TOP 10 hub genes were screened adopting the Cytoscape software and the gene expression was predicted or determined by the GEO and Gene Expression Profiling Interactive Analysis (GEPIA) databases or quantitative reverse transcription PCR (qRT-PCR) and western blot. The molecular docking was executed in the Autodock Vina software. Cellular functional assays (cell counting kit-8 (CCK-8), colony formation, 5-ethynyl-2′-deoxyuridine (EdU) staining, flow cytometry, wound healing, transwell, and tube formation) were conducted. Network pharmacology integrating the Gene Expression Omnibus (GEO), Comparative Toxicogenomics Database (CTD), SwissTargetPrediction, and GeneCards databases identified potential Cu-B targets. Protein-protein interaction (PPI) network (String), Gene Ontology/Kyoto Encyclopedia of Genes and Genomes (GO/KEGG) enrichment analysis (DAVID), and molecular docking (Autodock Vina) were performed. Hub gene validation employed quantitative reverse transcription PCR (qRT-PCR), western blot, and rescue experiments. Cu-B dose-dependently restrained the malignant behaviors of FaDu cells. Bioinformatics analysis prioritized transforming growth factor beta 1 (TGFB1) as a core target, confirmed by its overexpression in hypopharyngeal cancer. Molecular docking revealed strong Cu-B-TGFB1 binding. Meanwhile, Cu-B retarded FaDu cell malignant behaviors by repressing TGFB1 expression. The network pharmacology along with bioinformatics analysis and molecular docking confirm that Cu-B may combine with TGFB1 to confine the malignant progression of hypopharyngeal cancer.