Novel insights into the anticancer activity of clinically derived Colicin N and its association with EMT-related molecular modulation: An integrated computational and experimental study.

Epithelial-mesenchymal transition (EMT) promotes cancer progression and metastasis. Here, clinically isolated, UTI-derived Colicin N was evaluated for anticancer activity and EMT-associated molecular modulation. Among 179 UTI-derived E. coli isolates, 8 were colN-positive; purified Colicin N showed 97.48% RP-HPLC purity and a molecular mass of 41.74 kDa. Docking and molecular dynamics predicted stable Colicin N interactions with CDH1/CDH2, with wild-type complexes showing up to 4-5 intermolecular hydrogen bonds and larger interfacial contact areas than N280A and K276E complexes. Colicin N reduced cancer-cell viability to 45-60% and increased apoptosis to 50-55% at 25 μM, while ADF viability remained 75-80%. Migration decreased to 28% in A549 and 19.03% in U87 at 24 h. PCR and Western blot demonstrated CDH1/CDH2 modulation, with altered TGF-β/SMAD and PI3K/AKT/mTOR-associated genes. Overall, UTI derived clinical Colicin N showed anticancer activity associated with modulation of EMT-related molecular markers and signalling pathways.