Development of an extracellular vesicle-based shRNA delivery system to overcome cisplatin resistance in non-small cell lung cancer by targeting STAT3
Makaleler > Tam Makale · 2026
Yazarlar: Mustafa KOTMAKÇI, Vildan BOZOK
Ege Tıp Dergisi, Uluslararası, 2026, cilt 65, ss 507-518
DOI: 10.19161/etd.1952225
Özet
Aim: Cisplatin resistance remains a major clinical challenge in non-small cell lung cancer, frequently driven by STAT3 activation. This study aimed to engineer an autologous extracellular vesicle (EV) formulation loaded with anti-STAT3 shRNA-encoding plasmid DNA to resensitize cisplatin-resistant Calu1 lung cancer cells.Materials and Methods: EVs were isolated from parental Calu1 cell culture supernatants via modified differential ultracentrifugation and characterized using electron microscopy, atomic force microscopy, and dynamic light scattering. Plasmid DNA was loaded into EVs via lipofection-mediated membrane fusion. Formulation integrity and protection were assessed using agarose gel electrophoresis and a DNase I assay. In vitro cytotoxicity, STAT3 knockdown, and cisplatin resensitization were evaluated in a cisplatin-resistant subline (CR-Calu1) using WST-1 viability assays, RT-qPCR, and Western blot analysis.Results: Isolated EVs displayed characteristic spherical morphology and a typical size distribution. The biomimetic formulation successfully encapsulated the pDNA, conferring robust protection against DNase I degradation. In CR-Calu1 cells, pshSTAT3-loaded EVs significantly downregulated STAT3 mRNA expression (48–58% reduction) and correspondingly decreased protein levels. Crucially, combining these EVs with sub-lethal cisplatin doses synergistically reduced cell viability compared to cisplatin alone, effectively restoring chemosensitivity. Stability optimization identified 7.5% glycine as the superior cryoprotectant, maintaining EV structural integrity post-lyophilization.Conclusion: We successfully engineered a stable, lyophilizable EV-based gene delivery platform. Targeting STAT3 via this autologous system effectively reverses acquired drug resistance, representing a viable therapeutic strategy for refractory NSCLC.