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Empagliflozin Modulates Apoptosis and Autophagy Pathways in Lung Cancer Through Network Pharmacology and In Vitro Analysis

Year: 2026

Type: article

Abstract: ABSTRACT Lung cancer remains a major cause of cancer‐related mortality, highlighting the need for novel therapeutic strategies. Empagliflozin (EMPA), a sodium–glucose cotransporter 2 inhibitor, has demonstrated emerging anticancer activity; however, its effects on apoptosis–autophagy crosstalk in lung cancer remain incompletely understood. This study investigated the anticancer effects of EMPA and the influence of hydroxychloroquine (HCQ)‐mediated autophagy inhibition in A549 and HOP‐92 human lung cancer cells. Network pharmacology with explicit database association‐score cutoffs, and molecular docking validated by redocking of a co‐crystallized reference ligand, were used to identify potential EMPA‐associated targets and pathways. Cytotoxicity of EMPA, HCQ, and their combination was evaluated by MTT assay, followed by combination index and Bliss analyses. Cell‐cycle distribution and apoptosis were assessed by flow cytometry. Expression of apoptosis‐ and autophagy‐related markers was evaluated by RT‐qPCR, cleaved caspase‐3 analysis, and Western blotting. Network analysis identified 239 overlapping EMPA–lung cancer targets, of which 46 were retained after applying an Open Targets association‐score cutoff of 0.30. Docking, validated by reproducing the erlotinib crystal pose in the EGFR kinase domain (heavy‐atom RMSD 1.30 ± 0.02 Å), indicated that EMPA can be accommodated at surface sites of ANXA8 (−7.6 kcal/mol) and the p62/SQSTM1 ZZ domain (−7.0 kcal/mol) and within the ligand‐binding pocket of PPARγ (−7.5 kcal/mol; PDB 8BF1, the PPARγ–PGC‐1α coactivator complex). EMPA reduced cell viability in both cell lines, with greater sensitivity in A549 cells. Combining EMPA with HCQ markedly reduced IC 50 values and demonstrated synergistic interactions at IC 50 . EMPA altered cell‐cycle progression and increased apoptotic cell death, accompanied by enhanced BAX and CASP3 expression and cleaved caspase‐3 activation. EMPA increased LC3‐II without p62 accumulation, consistent with increased autophagic activity, whereas HCQ‐containing treatments induced LC3‐II and p62 accumulation and enhanced apoptotic signaling. EMPA exerts antiproliferative and pro‐apoptotic effects in lung cancer cells while modulating autophagy. Inhibition of lysosomal autophagic degradation by HCQ enhances EMPA‐mediated cytotoxicity and apoptosis, supporting further investigation of this combination as an anticancer strategy.

Source: Journal of Biochemical and Molecular Toxicology

DOI: 10.1002/jbt.71174