Abstract <p>This study aimed to clarify how miR-101-3p regulates cisplatin (DDP) resistance in non-small cell lung cancer (NSCLC). Candidate targets of miR-101-3p were first identified through bioinformatics analysis. miR-101-3p expression was compared between DDP-sensitive and DDP-resistant NSCLC cell lines. Functional assays assessed the effects of miR-101-3p overexpression on cell behavior and drug sensitivity. Dual-luciferase reporter assays were used to confirm direct binding of miR-101-3p to YWHAZ, while rescue experiments assessed the role of YWHAZ in mediating these effects. YWHAZ was identified as a direct target of miR-101-3p and was found to be upregulated in DDP-resistant NSCLC cells. Overexpression of miR-101-3p suppressed proliferation, invasion, and migration, while promoting apoptosis and lowering the DDP IC50. These effects were reversed by YWHAZ overexpression, demonstrating its involvement in DDP resistance. Overall, miR-101-3p reduces DDP resistance in NSCLC by targeting YWHAZ, thereby inhibiting malignant behaviors and restoring chemosensitivity. These findings suggest miR-101-3p as a promising therapeutic target for overcoming DDP resistance in NSCLC.</p>