Hesperadin sensitizes gastric cancer cells to cisplatin via NOX1-dependent oxidative stress.

The therapeutic efficacy of cisplatin in gastric cancer (GC) is frequently constrained by chemoresistance and systemic toxicity. Targeted small molecules hold promise for providing synergistic effects that enhance treatment outcomes. GC cell viability was assessed via a Cell Counting Kit-8 (CCK-8) assay. Apoptosis, intracellular reactive oxygen species (ROS) levels, and the mitochondrial membrane potential were evaluated via flow cytometry and JC-1 staining. Transcriptomic analysis [RNA-sequencing (RNA-Seq)] was subsequently conducted to identify differentially regulated pathways, followed by validation via reverse transcription-quantitative PCR and western blotting. DNA damage was measured by γ-H2AX immunofluorescence, and clonogenic survival was examined. The NOX1-specific inhibitor ML171 was used to verify its mechanistic involvement. Hesperadin markedly suppressed GC cell proliferation in a dose-dependent manner and induced mitochondrial apoptosis by regulating the Bcl-2/Bax ratio and activating caspase pathways. RNA-Seq analysis revealed significant upregulation of NOX1 and activation of oxidative stress-associated pathways following Hesperadin treatment. Cotreatment with Hesperadin and cisplatin markedly reduced the IC of cisplatin, increased ROS accumulation, aggravated DNA damage, and potentiated apoptotic cell death. Notably, ML171 attenuated ROS generation and apoptotic effects, confirming the occurrence of a NOX1-dependent mechanism. In conclusion, Hesperadin enhances the sensitivity of GC cells to cisplatin by inducing NOX1-mediated oxidative stress and promoting mitochondrial dysfunction-driven apoptosis, underscoring its potential as an effective combinatorial therapeutic strategy for GC treatment.