Identification and validation of adenylate uridylate- (AU-) rich element (ARE)-related biomarkers in the process of treating lung cancer mice with Wenyang Hualiu Tang through bulk RNA sequencing and animal model

Lung cancer (LC) remains the leading cause of cancer-related mortality. While the traditional formula Wenyang Hualiu Tang (WYHLT) has shown efficacy against LC, the role of adenylate uridylate (AU)-rich element (ARE)-related genes (AREGs) in this disease and its treatment is unexplored. This study aimed to elucidate the mechanism by which AREGs are involved in WYHLT treatment of LC. Transcriptome sequencing of a mouse LC model with WYHLT intervention (low, medium, high-dose) identified four key biomarkers: GREB1, MUC2, NPTX2, and PPIP5K1. Enrichment analysis linked MUC2, NPTX2, and PPIP5K1 to translation. A constructed regulatory network revealed interactions involving three biomarkers (PPIP5K1, GREB1, NPTX2), 16 long non-coding RNAs (lncRNAs), and 10 circular RNAs (circRNAs), with lncRNA Malat1 targeting four microRNAs (miRNAs). Drug prediction analysis identified seven drugs targeting MUC2 and one targeting NPTX2, with PD-98059 showing the highest binding affinity for MUC2. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) validation confirmed that WYHLT intervention increased PPIP5K1 and MUC2 expression and decreased GREB1 and NPTX2 expression compared to the LC model. This study identifies GREB1, MUC2, NPTX2, and PPIP5K1 as potential biomarkers involved in WYHLT treatment of LC. The findings provide new insights into the molecular mechanisms of WYHLT and novel directions for LC therapeutic research.