CAF-derived exosomal LncRNA ANRIL promotes glycolytic metabolism and proliferation in non-small cell lung cancer via the miR-186-5p/HIF-1α axis.

BACKGROUND

Cancer-associated fibroblasts (CAFs) critically regulate tumor microenvironment remodeling, with exosomes (Exos) derived from CAFs serving as key mediators of intercellular communication. However, the functional significance of CAF-derived exosomal long non-coding RNA ANRIL (lncRNA ANRIL) in modulating non-small cell lung cancer (NSCLC) glycolytic metabolism and proliferation remains incompletely characterized. This study systematically investigated the lncRNA ANRIL/miR-186-5p/HIF-1α regulatory axis in NSCLC progression.

METHODS

CAFs were isolated from fresh NSCLC surgical specimens, followed by Exo isolation through differential ultracentrifugation. The functional impacts of CAF-derived Exos on NSCLC cellular proliferation and glycolytic activity were comprehensively evaluated using CCK-8 assays, EdU incorporation tests, glucose consumption measurements, lactate production quantification, and extracellular acidification rate (ECAR) assessments. Molecular interactions within the ANRIL/miR-186-5p/HIF-1α axis were mechanistically validated through dual-luciferase reporter systems, RNA pull-down assays, and RT-qPCR profiling. Functional validation was achieved via targeted overexpression and siRNA-mediated knockdown approaches.

RESULTS

CAF-derived Exos significantly enhanced NSCLC cell proliferation indices and glycolytic parameters. CRISPR-mediated ANRIL silencing in CAFs substantially attenuated these pro-tumorigenic effects, establishing exosomal ANRIL as a critical molecular effector. Integrated bioinformatics prediction and experimental validation confirmed direct binding between lncRNA ANRIL and miR-186-5p, with subsequent identification of HIF-1α as the downstream target of miR-186-5p. Functional perturbation experiments demonstrated that miR-186-5p overexpression or HIF-1α knockdown effectively suppressed NSCLC proliferation and glycolysis, while rescue assays confirmed HIF-1α as the terminal mediator of miR-186-5p regulatory effects.

CONCLUSIONS

This mechanistic study demonstrates that CAF-secreted exosomal lncRNA ANRIL drives NSCLC progression by enhancing glycolytic metabolism through competitive sponging of miR-186-5p, thereby derepressing HIF-1α expression. Our findings provide novel insights into exosome-mediated metabolic reprogramming in NSCLC and propose therapeutic targeting of the ANRIL/miR-186-5p/HIF-1α signaling axis as a potential precision medicine strategy.