© 2022 Elsevier B.V.Bacterial membrane vesicles have recently attracted attention as drug delivery systems for antineoplastic agents to target cancerous cells and reduce the side effects of free cytostatic drugs. Due to the complex nature of interactions between the gut microbiota and the host, vesicles released by commensal strains are thought to play a key role in signaling processes in the intestinal mucosa. Herein, it was aimed to investigate the cytotoxic and anti-apoptotic effects of vesicles released from Enterococcus faecalis that the most abundant commensal bacteria in the intestinal mucosa on HT-29 colon cancer cells. In this context, naturally released nano-sized membrane vesicles were successfully isolated and loaded with the antineoplastic drug Capecitabine. The cytotoxic and anti-proliferative effects of drug-loaded membrane vesicles on colon cancer cells have been demonstrated by XTT and the live/dead staining assay. Furthermore, mRNA expression analyses demonstrated that the expressions of TP53 and Caspase-3 were upregulated, confirming that apoptosis was enhanced in cancer cells upon treatment with drug-loaded vesicles. These findings suggest that membrane vesicles have intriguing potentials as living-derived nano-drug delivery systems, particularly for chemotherapeutic drugs.