ENO1 regulates trastuzumab resistance in gastric cancer cells through IRP1

This study aims to investigate the metabolic alterations, their impacts on trastuzumab resistance, and potential therapeutic strategies to overcome resistance to trastuzumab in HER2-positive gastric cancer. Single-cell RNA sequencing (scRNA-seq) was employed to classify gastric cancer cells, and glycolysis-related genes, particularly Enolase 1 (ENO1), were quantified by RT-PCR and Western blotting. ENO1 overexpression and knockdown models were developed to explore its role in trastuzumab resistance. RNA immunoprecipitation (RIP) assays explored ENO1's interaction with Iron Regulatory Protein 1 (IRP1). Additionally, the expression of ferroptosis-related genes was analyzed to determine their association with trastuzumab sensitivity. Our study revealed a significant upregulation of glycolysis in trastuzumab-resistant gastric cancer cells, primarily driven by the overexpression of the glycolytic enzyme ENO1. This metabolic reprogramming was identified as a pivotal mechanism underlying trastuzumab resistance, contributing to enhanced cell survival, increased colony formation, and accelerated tumor growth. Detailed analyses demonstrated that ENO1 overexpression not only facilitated glycolysis but also inhibited the expression of the tricarboxylic acid (TCA) cycle-related gene IRP1. This inhibition modulated ferroptosis, and further promoted resistance. The dual role of ENO1 in regulating both glycolysis and ferroptosis underscored its critical involvement in metabolic adaptations. Metabolic reprogramming, particularly through ENO1, plays a key role in trastuzumab resistance. Targeting glycolysis gene ENO1, along with elucidating the regulatory mechanisms of IRP1 and ferroptosis, presents promising therapeutic strategies to overcome trastuzumab resistance in gastric cancer.