NRP1 silencing induces ROS-mediated cell cycle arrest and mitochondrial apoptosis in non-small cell lung cancer via upregulation of AGER.

OBJECTIVE

The specific role of Neuropilin-1 (NRP1) and its link to oxidative stress in Non-Small Cell Lung Cancer (NSCLC) progression remains unclear.

METHODS

Integrated bioinformatics, including weighted gene co-expression network analysis and machine learning, were applied to the TCGA cohort and validated in an age-stratified clinical cohort. In vitro assays evaluated proliferation, apoptosis, ATP, and Reactive Oxygen Species (ROS) in NRP1-silenced H1299 cells. Mechanisms were investigated using endogenous Co-Immunoprecipitation (Co-IP) and rescue experiments with AGER siRNA and the ROS scavenger N-Acetylcysteine (NAC).

RESULTS

Bioinformatics identified AGER as a key NRP1 downstream target. Clinically, NRP1 was significantly upregulated exclusively in elderly (≥65-years) NSCLC patients, whereas AGER was consistently downregulated. Functionally, NRP1 silencing inhibited proliferation, induced G1-phase arrest, and triggered mitochondria-dependent apoptosis characterized by severe ATP depletion and ROS accumulation. Endogenous Co-IP confirmed a direct physical interaction between NRP1 and AGER. Importantly, the oxidative stress and apoptotic phenotypes induced by NRP1 knockdown were effectively reversed by AGER silencing or NAC treatment.

CONCLUSION

The NRP1-AGER-ROS axis critically regulates tumor progression and oxidative stress in NSCLC, particularly in elderly patients, representing a promising age-associated therapeutic target.