Mitochondrial uncoupler BAM15 attenuates cell proliferation and tumor growth in non-small cell lung cancer

Lung cancer remains the leading cause of cancer-related mortality worldwide, with non-small cell lung cancer (NSCLC) accounting for 80–85% of all cases, highlighting the urgent need for novel therapeutic strategies. BAM15, a mitochondria-targeted uncoupler, has demonstrated therapeutic potential in metabolic disorders and several cancer types; however, its role in NSCLC progression remains poorly understood. This study aimed to evaluate the antitumor effects of BAM15 in human NSCLC cells (A549, H1299) and elucidate the underlying mechanisms, with in vivo validation. The results showed that BAM15 treatment dose-dependently inhibited the viability of NSCLC cells (IC50: 4.013 µM for A549, 7.897 µM for H1299) and suppressed their colony formation, migration and invasion. Furthermore, BAM15 not only induced G1/G0 phase arrest but also apoptosis in NSCLC cells. RNA sequencing identified 2,270 differentially expressed genes (DEGs) in response to BAM15 treatment. Pathway analysis showed that BAM15 downregulated cell signaling pathways involved in DNA replication and cell cycle, and upregulated those associated with inflammatory response (e.g., TNF, IL-17, NF-κB signaling) and MAPK/PI3K-Akt signaling. Western blot confirmed that BAM15 downregulated key cell cycle regulators, including CDK2, CDK6, and PCNA. In vivo experiment, BAM15 administration significantly reduced xenograft tumor weight and volume and decreased the number of PCNA-positive tumor cells. In conclusion, BAM15 exerts potent antitumor effects against NSCLC both in vitro and in vivo by disrupting DNA replication and cell cycle progression, likely through modulation of cell cycle regulators and downstream signaling pathways. These findings suggest BAM15 as a promising candidate for targeted NSCLC therapy.