BACKGROUND

Gastric cancer (GC) remains a leading cause of cancer-related mortality worldwide due to late-stage diagnosis and therapeutic resistance. Oxidative stress and autophagy play paradoxical roles in GC progression, yet their regulatory mechanisms remain incompletely understood. This study investigates the molecular interplay between chemokine CCL2, oxidative stress, autophagy, and m6A RNA demethylase fat mass and Obesity-associated protein (FTO) in GC pathogenesis.

METHODS

Oxidative stress-related genes were identified through bioinformatics analysis using the GeneCards and GEO databases. Key hub genes were determined via protein-protein interaction network analysis. The expression of CCL2 and FTO in GC tissues and cell lines was examined by quantitative real-time PCR (qRT-PCR), western blotting, and enzyme-linked immunosorbent assay (ELISA). Functional assays, including colony formation, apoptosis analysis, reactive oxygen species (ROS) measurement, and autophagy detection, were performed in GC cell lines. The m6A regulatory effects of FTO on CCL2 were evaluated using RNA immunoprecipitation (RIP), methylated RNA immunoprecipitation (MeRIP), and RNA stability assays. Furthermore, a GC xenograft model was used to validate in vivo tumorigenic effects.

RESULTS

Bioinformatics analyses identified significant dysregulation of oxidative stress-associated genes, notably highlighting CCL2 as a central hub gene strongly linked to poor prognosis. Experimental validation demonstrated elevated expression of CCL2 in GC tissues, patient serum, and cell lines, correlating with reduced overall survival. Functional assays revealed that CCL2 promotes GC cell proliferation and survival by suppressing autophagy and attenuating oxidative stress-induced apoptosis. Mechanistically, we identified that FTO-mediated m6A demethylation enhances CCL2 mRNA stability and expression, thus facilitating GC progression. Notably, FTO knockdown resulted in increased oxidative stress, enhanced autophagy, and reduced tumorigenicity, which were reversed by CCL2 overexpression. Moreover, YTHDF2, an m6A reader protein, was identified as a negative regulator of CCL2, promoting its degradation and counteracting FTO-mediated stabilization. Additionally, xenograft experiments confirmed the tumor-promoting effect of the FTO-CCL2 axis in vivo.

CONCLUSIONS

This study uncovers a novel regulatory mechanism by which FTO-mediated m6A demethylation stabilizes CCL2, leading to oxidative stress reduction, autophagy inhibition, and enhanced GC progression. Targeting the FTO-CCL2 axis may represent a promising therapeutic strategy for GC treatment.