<bold>Background:</bold> <italic>Staphylococcus aureus</italic> (SA) and cigarette smoke exposure in asthmatics are associated with asthma severity and airway remodeling. However, their interaction mechanisms and impacts remain unclear. This study investigated transcriptomic changes in human bronchial epithelial cells exposed to cigarette smoke extract (CSE), SA culture supernatant, and their co-exposure, aiming to elucidate the synergistic effects on airways. <bold>Methods:</bold> BEAS-2B cells were exposed to CSE, SA culture supernatant, or their co-exposure for 24 hours. RNA was extracted, and RNA sequencing (RNA-Seq) was performed. Differentially expressed genes (DEGs), Gene Ontology (GO) enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway, and gene-gene interaction (GGI) network analysis were conducted. Quantitative RT-PCR (qRT-PCR) validated the RNA-Seq findings. <bold>Results:</bold> RNA-Seq revealed distinct gene expression profiles across treatment groups, with the co-exposure showing significantly greater number of DEGs compared to individual exposures. GO and KEGG pathway analyses highlighted enrichment of pathways related to inflammation, cellular stress, and tissue remodeling in the co-exposure group. GGI network analysis suggested involvement in neutrophil migration, IL-17 production, epithelial cell adhesion/apoptosis, and tissue remodeling. qRT-PCR confirmed synergistic upregulation of key DEGs in the co-exposure group. <bold>Conclusion:</bold> These findings demonstrate synergistic inflammatory effects of SA and CSE co-exposure on airway epithelial cells, potentially providing clues to their interaction mechanisms in airway inflammation and remodeling pathogenesis.