NT5E Mediates Cisplatin Resistance in NSCLC Through the PI3K‐AKT Signaling Pathway

Objective: The aim of this study was to investigate the role of NT5E and its mechanism in cisplatin (diamminedichloroplatinum, DDP) resistance in non‐small cell lung cancer (NSCLC) and to explore the value of NT5E in the clinical prognosis of NSCLC patients. Methods: First, the genes related to drug resistance were identified by constructing an A549/DDP cell line with DDP resistance and performing transcriptome sequencing and bioinformatics analysis. Gene ontology (GO) as well as the Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathways were enriched for the related genes using the Metascape database. The STRING database was further applied to construct protein–protein interaction networks (PPIs), and heatmaps were used to visualize the coexpression relationships of genes in the modules, as well as to screen for key genes. Subsequently, immunohistochemistry and molecular biology experiments were conducted to confirm the expression levels of NT5E in DDP‐resistant NSCLC tissues and cells. Additionally, its effects on cell proliferation, migration, and invasion were assessed through NT5E knockdown assays. Finally, RNA sequencing and signaling pathway analyses revealed that NT5E regulates DDP resistance in NSCLC cells through the PI3K‐AKT signaling pathway and analyzed the correlation between the expression of NT5E and survival outcomes in NSCLC patients. Results: NT5E was found to be upregulated in drug‐resistant cells and increased in DDP‐resistant NSCLC tissues and cells. The drug‐resistant genes were primarily involved in cell adhesion and the extracellular matrix. Functional experiments confirmed that the knockdown of NT5E inhibited cell proliferation, migration, and invasion. Signaling pathway analysis indicated that NT5E mediated the resistance of NSCLC cells to DDP through the PI3K/AKT signaling pathway. Moreover, high NT5E expression was found to be negatively correlated with the overall survival of NSCLC patients. Conclusion: NT5E may be a key gene for DDP resistance in NSCLC cells, while its high expression may indicate a poor prognosis for patients. This finding provides important insights for a deeper understanding of the mechanism of drug resistance in lung cancer and serves as a theoretical basis for the development of new therapeutic strategies and drugs.