Background <p>This study aims to examine the potential value of lncRNA PAX8-AS1 and miR-96-5p as diagnostic markers in non-small cell lung cancer (NSCLC). The goal is to provide a reference for improving adverse outcomes in NSCLC and developing effective early interventions in the clinic.</p> Methods <p>A total of 112 NSCLC patients and 70 healthy volunteers were recruited as the study subjects. The expression levels of PAX8-AS1 and miR-96-5p were measured using fluorescence quantitative PCR. The expression levels of PAX8-AS1 and miR-96-5p were detected by fluorescence quantitative PCR. A dual luciferase reporter assay was used to detect the binding of PAX8-AS1 and miR-96-5p. ROC, Kaplan-Meier, and Cox regression analysis was used to assess the diagnostic and prognostic value of PAX8-AS1 and miR-96-5p in NSCLC.</p> Results <p>In NSCLC, PAX8-AS1 was found to be downregulated, whereas miR-96-5p was upregulated. These two molecules exhibited a negative correlation and a target-binding relationship. Receiver operating characteristic (ROC) curve analysis demonstrated that PAX8-AS1 and miR-96-5p had practical diagnostic values for distinguishing NSCLC patients. Mortality increased in NSCLC patients with low PAX8-AS1 levels and decreased in those with low miR-96-5p levels. Cox regression analysis showed that PAX8-AS1 and miR-96-5p could serve as independent predictors of prognostic survival in NSCLC patients.</p> Conclusion <p>PAX8-AS1 and miR-96-5p can serve as potential diagnostic biomarkers for NSCLC patients.</p>