Mechanism of Non–Small Cell Lung Cancer–Derived Extracellular Vesicle miRNA hsa-let-7b-5p Targeting AP1S1 to Regulate M2 Macrophage Polarization

Abstract

Background

Non–small cell lung cancer (NSCLC) accounts for over 80% of lung cancer cases. Further, the complex tumor immune microenvironment (TIME) is a critical factor in treatment resistance and poor prognosis associated with tumors. Tumor-associated macrophages (TAMs), a major component of the TIME, significantly promote tumor progression through their polarization toward the immunosuppressive M2 phenotype. Reportedly, NSCLC cells regulate TAM polarization by secreting extracellular vesicles (EVs) to deliver miRNAs; however, the specific underlying molecular mechanisms remain unclear. In this study, we aimed to elucidate the regulatory role of miRNAs derived from NSCLC EVs in TAM polarization and explore potential novel therapeutic targets.

Methods

Through high-throughput sequencing and bioinformatics analysis, key regulatory targets were screened. Ki-67 staining was employed to detect cell proliferation, flow cytometry was performed to analyze cell apoptosis, RT-qPCR and Western blot were used to measure mRNA and protein expression levels, and Transwell assays were conducted to assess cell migration and invasion capabilities to investigate the molecular mechanisms underlying the miRNA-mediated regulation of TAM polarization by NSCLC-derived EVs.

Results

NSCLC-derived EVs were successfully isolated and characterized. Bioinformatics analysis of EVs′ miRNA sequencing data revealed that the hsa-let-7b-5p/Adaptor-Related Protein Complex 1 subunit sigma 1 (AP1S1) axis may be a key regulator of TAM polarization. In vitro experiments confirmed that the hsa-let-7b-5p mimic potentially suppressed M2 polarization of TAMs via the AP1S1/p53 signaling axis, thereby attenuating the proliferation, migration, and invasion capabilities of NSCLC cells.

Conclusion

This study revealed the molecular mechanism by which hsa-let-7b-5p reshapes the immune microenvironment of NSCLC cells by targeting and inhibiting AP1S1 expression, thereby regulating the polarization of TAMs toward the M2 phenotype. Thus, the hsa-let-7b-5p/AP1S1 axis may serve as a potential therapeutic target for NSCLC immunotherapy, providing novel strategies for improving patient prognosis.