Hybrid liposomes (HLs) composed of vesicular and micellar surfactants have inhibitory effects on the growth of tumor cells in vitro and in vivo. Successful clinical chemotherapy with drug-free HLs to patient with lymphoma has been reported after approval by the Committe of Bioethics. However, the therapeutic effects of HLs on the metastasis of colon carcinoma cells have not yet been elucidated. In this study, the therapeutic effects of HLs composed of L-alpha-dimyristoylphosphatidylcholine (DMPC) and polyoxyethylene (23) dodecyl ether [C(12)(EO)(23)] on the metastasis of colon carcinoma (Colon26) cells were examined in vivo. Marked high therapeutic effects were obtained in the hepatic metastasis mice model after the treatment with HLs. Furthermore, optical microscopic analysis indicated that HLs could induce the apoptosis of colon carcinoma cells in vivo. No toxicity was observed in the hepatic metastasis mice model after intravenously injecting HLs. Therapeutic effects along with the induction of apoptosis by HLs without any drugs on hepatic metastasis were revealed on the basis of optical microscopic analysis for the first time in vivo.