Investigation of the effects of nitrofen on global gene expression in a lung cell line using the RNA-Seq method

Aim: This study aimed to evaluate the molecular effects of nitrofen, an environmental toxicant, on the human bronchial epithelial cell line BEAS-2B at the transcriptomic level and to validate the findings with complementary methods. Materials and Methods: BEAS-2B cells were cultured and treated with 800 µM nitrofen for 24 hours, as determined by the cell viability assay. Total RNA was isolated and sequenced for RNA-Seq analysis. Differential gene expression analysis was performed, and significant genes were assessed by GO and KEGG enrichment analyses. Selected genes were validated by RT-qPCR, and protein-level changes were examined by Western blot. Results: RNA-Seq analysis identified 502 significantly differentially expressed genes, including 320 upregulated and 182 downregulated genes. Enrichment analyses showed that nitrofen exposure affected pathways related to cellular stress response, metabolic processes, and epithelial function. RT-qPCR largely confirmed the RNA-Seq findings, and Western blot analysis showed similar trends at the protein level for selected targets. Conclusion: Nitrofen exposure induced extensive gene expression changes in BEAS-2B cells and affected pathways associated with stress response, metabolic regulation, and epithelial integrity. These findings suggest that nitrofen exerts multifaceted effects on human airway epithelium. Keywords: BEAS-2B, Differential Gene Expression, Nitrofen, RNA-Seq, Toxicity