BAY-876 restores lenvatinib sensitivity in resistant hepatocellular carcinoma cells by disrupting the GLUT1-SLC7A11-NADPH redox axis.

BACKGROUND

Acquired resistance limits the efficacy of lenvatinib in hepatocellular carcinoma (HCC). This study evaluated whether GLUT1 inhibition by BAY-876 reverses lenvatinib resistance by disrupting SLC7A11-associated redox buffering.

METHODS

Lenvatinib-resistant HuH-7-LR and Hep 3B-LR cells were generated by stepwise drug exposure. Drug sensitivity, combination synergy, glucose uptake, colony formation, migration, ATP, NADPH/NADP⁺ and GSH/GSSG ratios, intracellular thiols, ROS, apoptosis, gene and protein expression, N-acetyl-L-cysteine (NAC) rescue, and SLC7A11 knockdown were evaluated.

RESULTS

Resistant cells exhibited increased lenvatinib IC₅₀ values, glucose uptake, and GLUT1/SLC7A11 expression. BAY-876 preferentially inhibited glucose uptake and viability in resistant cells and restored lenvatinib sensitivity to near-parental levels. Combined treatment produced synergistic growth inhibition, reduced colony formation and migration, depleted ATP and NADPH, disrupted GSH/GSSG balance, decreased intracellular thiols, and increased ROS. It also induced apoptosis, evidenced by Annexin V positivity, caspase-3/7 activation, BAX induction, BCL2 suppression, and PARP/caspase-3 cleavage. NAC partially restored viability and redox balance. SLC7A11 knockdown reduced BAY-876-associated thiol depletion and ROS accumulation, supporting an SLC7A11-dependent redox component.

CONCLUSION

BAY-876 resensitized lenvatinib-resistant HCC cells by restricting glucose uptake, disrupting NADPH-dependent redox buffering, and promoting redox stress-induced apoptosis. These findings support the GLUT1-SLC7A11-NADPH axis as an in vitro therapeutic vulnerability, while definitive evidence of disulfidptosis requires further investigation.