Modulation of almonertinib resistance in non-small cell lung cancer by cancer-associated fibroblasts through HK2-mediated glycolysis and SKP2 signaling

BackgroundAlmonertinib is a third-generation EGFR-TKI, and studies on its resistance mechanisms are lacking. Cancer-associated fibroblasts (CAFs) can influence resistance to targeted therapeutics, but their role and mechanism of action in relation to almonertinib resistance are unclear. The study explored relationships among glycolysis, cancer-associated fibroblasts (CAFs), and almonertinib resistance.MethodsA dose-escalation method was used to develop the almonertinib-resistant cell line H1975AR. Hexokinase 2 (HK2) effects on almonertinib resistance were evaluated using Cell Counting Kit-8 assays, transcriptome sequencing, western blotting, real-time PCR, siRNA assays, glucose consumption, and lactate production assays. Differential gene expression analysis and siRNA assays in H1975 cells cultured with CAF-conditioned medium (H1975/CAF-CM) revealed S-phase kinase-associated protein 2 (SKP2) as a target driving the effects of HK2. The impact of almonertinib and HK2 inhibitors on H1975/CAF-CM cells was assessed using colony formation, wound healing, transwell, and flow cytometry apoptosis assays.ResultsH1975AR cells displayed elevated glycolysis and HK2. Subsequently, we showed that knockdown reduced almonertinib resistance in cells. H1975/CAF-CM induced almonertinib resistance and upregulation of HK2, which was reversed by knockdown of SKP2. CAFs regulate HK2-mediated glycolysis through SKP2, promoting almonertinib resistance in NSCLC.ConclusionsCAFs regulate HK2-mediated glycolysis through SKP2, which promotes almonertinib resistance in NSCLC.