HPV VLPs-based RNA-delivery system for synergistic BCL-2 silencing and mitochondrial apoptosis in NSCLC cells

• A tetra-functional assembly, siRNA_D-pep@VLP-SP5-2, was integrated in this study. • It induced targeting precision of NSCLC cell and silenced BCL-2 expression. • It enhanced Cyt c release and triggered mitochondrial apoptosis. • It achieved 91 % A549 apoptosis and 78.2 % tumor growth inhibition in mice model. Non-small cell lung cancer (NSCLC) resists apoptosis and lacks targeted therapies. We developed a tetra-functional assembly, siRNA_D-pep@VLP-SP5-2, integrating BCL-2 siRNA and a cationic mitochondrial destabilization D-peptide within the SP5-2-modified HPV virus-like particle (VLP). This multifunctional assembly integrate: (1) SP5-2-driven tumor targeting precision, (2) VLP-improved cellular uptake, (3) siRNA-induced BCL-2 silencing (70 % knockdown), and (4) D-peptide-triggered Cyt c release and mitochondrial apoptosis. In cells , siRNA_D-pep@VLP-SP5-2 induced 91 % apoptosis of A549 cells, while in A549 tumor-bearing mice, it reduced 78.2 % tumor growth without systemic toxicity. This multifunctional assembly pioneer’s co-delivery of gene and therapeutical peptide synergistically, overcoming key barriers in NSCLC precision therapy. In particular, the thoroughly revealed apoptotic kinetics for the RNAi therapeutics and peptide-mediated cytotoxicity, together with the modular design using VLP opens new avenues for multimodal cancer therapeutics, expecting to transition from “limit-target inhibition” toward “dynamic network-level regulation”, thereby establishing a paradigm-shifting framework for solid tumor treatment.