Dual Metabolic Inhibition by Berberine and Glutor Triggers AMPK/JNK-Dependent DNA Damage in Cancer Cells

Metabolic reprogramming is a key adaptive feature of malignant tumors and often leads to the failure of therapeutic strategies targeting a single metabolic pathway.In this study, we investigated the antitumor activity of combining berberine (BBR), a mitochondrial complex I inhibitor, with Glutor, a pan-glucose transporter inhibitor that blocks glucose uptake across multiple GLUT isoforms.The combination produced strong synergistic antiproliferative effects in HeLa, HepG2, and HCT116 cells, with Bliss synergy scores of 45.967, 34.219, and 26.972, respectively.Co-administration simultaneously suppressed glycolysis and oxidative phosphorylation (OXPHOS), resulting in severe ATP depletion and persistent activation of the AMPK signaling pathway, thereby inducing pronounced metabolic stress.Further mechanistic investigations revealed that such sustained metabolic stress effectively activated the JNK signaling pathway, which in turn exacerbated DNA damage, impaired homologous recombination repair, and ultimately triggered caspase-dependent apoptosis.Pharmacological inhibition or genetic silencing of JNK attenuated DNA damage and partially restored HR repair-related proteins, supporting JNK as a key functional mediator of the downstream stress response.In ex vivo tumor explant models, the combination also showed cooperative antitumor activity.These findings indicate that simultaneous restriction of glucose uptake/glycolysis and mitochondrial oxidative phosphorylation, termed "dual metabolic inhibition" cooperatively impairs tumor cell survival.The AMPK/JNK axis appears to be a critical mechanistic link underlying this antitumor effect.