Nitric oxide-enriched plasma-activated water synergistically enhances the anticancer activity of Bauhinia variegata via TC2N-mediated apoptotic responses in lung cancer cells

TC2N (Tac2-N-terminal expressed protein 1) is an oncogenic regulator implicated in lung cancer progression and represents a promising target for anticancer therapy. In this study, we investigated a novel plasma-assisted therapeutic strategy combining Bauhinia variegate ( B. variegate ) extract with nitric oxide-enriched plasma-activated water (NO-PAW) to enhance selective anticancer responses in lung cancer cells. Using H460 and A549 lung cancer cells along with normal MRC5 lung fibroblasts, we demonstrated that the combination treatment synergistically reduced cancer cell viability while exhibiting minimal cytotoxicity toward normal cells. Notably, the combined treatment reduced H460 cell viability by approximately 63%, whereas only minimal effects were observed in MRC5 cells, indicating high treatment selectivity. Mechanistically, the synergistic treatment enhanced membrane damage, intracellular ROS/RNS accumulation, γ-H2AX expression, and apoptotic signaling responses. Mechanistically, the synergistic treatment promoted TC2N suppression, enhanced Cdk5–p53 interactions, and activated apoptotic molecular signaling pathways. Molecular docking and 100 ns molecular simulations revealed stable interactions between major B. variegata bioactive compounds and TC2N-associated binding regions, both independently and in the presence of NO-derived reactive nitrogen species (NO, NO 2 , and NO 3 ). ADMET and bioavailability analyses further supported the therapeutic potential and safety profiles of the selected compounds. Collectively, these findings demonstrate that nitric oxide-enriched plasma-activated water synergistically enhances the anticancer efficacy of B. variegata through TC2N-mediated apoptotic responses, highlighting a promising plasma–botanical strategy for selective lung cancer therapy.