The identification of tumor-specific markers combined with targeted therapeutic approaches has yielded substantial clinical benefits in recent years. PIWI-interacting RNAs (piRNAs), which exert important influences on tumor development, hold considerable promise as targets for creating molecular agents against cancer. This investigation confirmed the capacity of piR-hsa-164586 to promote invasion and metastasis in non-small cell lung cancer (NSCLC) cells. Building upon earlier findings of its markedly elevated levels in NSCLC tissues and serum extracellular vesicles, the study employed both in vitro and in vivo models to substantiate this role. By integrating single-cell RNA sequencing (scRNA-seq) with supplementary molecular analyses, the work showed that elevated expression of piR-hsa-164586 modifies the tumor microenvironment in NSCLC through enrichment of mesenchymal features in tumor epithelial cells. In addition, piR-hsa-164586 participates in controlling the PI3K-AKT signaling cascade and epithelial–mesenchymal transition, which in turn accelerates the invasive and metastatic spread of NSCLC cells. The non-PIWI protein MYH9 interacts directly with piR-hsa-164586, and reduced expression of either molecule correlates with more positive outcomes in NSCLC. Overall, the findings delineate the precise mechanism through which piR-hsa-164586 functions as a tumor marker in NSCLC while offering critical support for the design of molecularly targeted treatments for this malignancy.
