Direct Fusion between Poly(ethylene oxide)-lipid Modified Liposomes and Murine Mitotic B16 Melanoma Cells

Abstract

The interactions between the poly(ethylene oxide)-bearing lipid (PEO-lipid) were investigated with the average number of ethylene oxide units of 5, 15 and 32-reconstituted egg PC liposomes and murine mitotic B16 melanoma cells. Water-soluble FITC-dextran (20kDa) and lipophilic octadecyl rhodamine B (OD-RhoB) were encapsulated in liposomes to study the interaction modes with these cells by fluorescence microscopic techniques. Both fluorescent probes loaded in the PEO-lipid (n 32, 20mol%)-reconstituted liposome were specifically transferred to the mitotic cells. This process was not inhibited at 4°C or after the treatment of endocytosis inhibitor cytochalasin B or D. Confocal fluorescence microscopic observation of the cells treated with the liposome at 4°C revealed that FITC-dextran and OD-RhoB were transferred to the cytosol and the plasma membrane, respectively. In addition, when the mitotic cells were treated with the PEO-lipid (n 32, 20mol%)-reconstituted liposome encapsulated diphtheria toxin fragment A (DTA), approximately 30% of the cells were killed by the DTA-dependent cytotoxicity. These data indicate that the PEO-lipid (n 32, 20mol%)-reconstituted liposome directly fused with the plasma membrane of the murine mitotic B16 melanoma cells.