Hyaluronic Acid-Ceramide-based Liposomes for Targeted Gene Delivery to CD44-Positive Cancer Cells

摘要

Hyaluronic acid-ceramide (HACE)-modified liposomes were designed using 1,2-dioleoyl-sn-glycero-3-phoshphoethanolamine (DOPE) and 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) for targeted delivery of therapeutic genes to the CD44 receptor-overexpressing cancer cells. Liposomes were prepared with different molar ratios of HACE; the most efficient formulation was tested for further in vitro experiments. The size and zeta potential of HACE-based liposomes were characterized by a Zetasizer. Lipoplex was then prepared at different nitrogen/phosphate (N/P) ratios; the gel retardation test showed strong DNA-binding affinity of liposomes for targeted gene delivery. Cytotoxicity of liposomes was evaluated by colorimetric assay (WST assay) for different cell lines such as MDA-MB-231 and NIH3T3 cells. HACE liposomes showed negligible cytotoxicity both in MDA-MB-231 and NIH3T3 cells that endow them for further therapeutic studies. We then examined the transfection efficiency of liposomes using luciferase reporter plasmid DNA. We found transfection efficiency of HACE-based liposomes was remarkably higher in case of MDA-MB-231 cells as compared to NIH3T3 cells. This result was indicative for higher receptor-binding endocytosis uptake of HACE liposomes and subsequent transfection in CD44 receptor-positive cell lines. Our findings have shown interesting prospective for tumor-targeted delivery of therapeutic gene in CD44 receptor-positive cells with less cytotoxic effects.