Resveratrol Modulates Lung Cancer-Associated Hub Genes Identified by Integrated Bioinformatics, Single-Cell Analysis, Molecular Docking, and Experimental Validation.

Abstract

Lung cancer remains a major cause of cancer-related mortality, and reliable molecular targets with therapeutic relevance are still needed. This study aimed to identify key genes associated with lung cancer progression and explore the potential regulatory effect of resveratrol. The GSE43458 dataset, including 80 lung cancer samples and 30 control samples, was used for differential expression analysis and weighted gene co-expression network analysis (WGCNA). Intersecting genes were subjected to protein-protein interaction network construction, Cytoscape-based hub gene screening, functional enrichment analysis, and immune infiltration analysis. Single-cell RNA sequencing data from GSE131907 were further analyzed to characterize cellular heterogeneity and hub gene distribution. Resveratrol was predicted as a candidate compound, followed by molecular docking with hub proteins. Finally, CCK-8, Western blotting, and qRT-PCR assays were performed in BEAS-2B and A549 cells. A total of 1,655 differentially expressed genes were identified, and WGCNA identified the 252-gene turquoise module as the module most strongly associated with the lung cancer phenotype (r = - 0.86, P = 6.0 × 10⁻³⁵). Intersection analysis yielded 250 candidate genes, representing 15.1% of all differentially expressed genes. Five hub genes, CASP3, DDX54, TP53BP1, CDKN2A, and ABT1, were identified and showed significantly increased expression in lung cancer tissues (P < 0.05). Functional enrichment analysis linked the candidate genes to apoptosis, cell-cycle regulation, DNA damage repair, immune responses, and cancer-related pathways, while immune infiltration and single-cell analyses revealed marked remodeling of immune and stromal components. Molecular docking predicted potential interactions between resveratrol and the five hub proteins, with the most favorable docking scores observed for TP53BP1 (- 7.0 kcal/mol) and DDX54 (- 6.9 kcal/mol). In vitro validation further showed that resveratrol treatment significantly reduced the mRNA and protein expression of the identified hub genes in A549 cells (P < 0.05). This study identified five lung cancer-associated hub genes and provided preliminary evidence that resveratrol may modulate their expression. These findings provide a multi-level molecular framework for further investigation of resveratrol-responsive networks in lung cancer.