MicroRNAs (miRNAs) are crucial in physiological and pathological processes and serve as biomarkers for various diseases. We previously validated seven miRNA biomarkers and nine in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). In this study, we observed distinct clustering patterns of LUAD or LUSC tissues compared to paired normal tissues based on miRNA expression levels, suggesting the potential involvement of circulating miRNAs in non-small-cell lung cancer (NSCLC) progression. To elucidate their biological function, we identified the most significant differentially expressed miRNAs (DE-miRNAs)-hsa-miR-451a, hsa-miR-139-5p and hsa-miR-126-5p-using The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) datasets. We then performed protein-protein interaction (PPI) analysis and constructed a miRNA-hub gene regulatory network based on targets predicted by several miRNA-target prediction tools. Additionally, we evaluated the biological functions of these miRNA biomarkers through EdU and wound healing assays in A549 cells. Our study identifies three miRNAs that may contribute to lung cancer progression by modulating cancer-related targets and highlights their potential as biomarkers. Future mechanistic investigations may provide novel insights into NSCLC pathogenesis and open new therapeutic avenues.