Abstract
BackgroundGene therapy has tremendous potential for both inherited and acquired diseases. However, delivery problems limited their clinical application, and new gene delivery vehicles with low cytotoxicity and high transfection efficiency are greatly required.MethodsIn this report, we designed and synthesized three amphiphilic molecules (L1–L3) with the structures involving 1, 4, 7, 10-tetraazacyclododecane (cyclen), imidazolium and a hydrophobic dodecyl chain. Their interactions with plasmid DNA were studied via electrophoretic gel retardation assays, fluorescent quenching experiments, dynamic light scattering and transmission electron microscopy. The in vitro gene transfection assay and cytotoxicity assay were conducted in four cell lines.ResultsResults indicated that L1 and L3-formed liposomes could effectively bind to DNA to form well-shaped nanoparticles. Combining with neutral lipid DOPE, L3 was found with high efficiency in gene transfer in three tumor cell lines including A549, HepG2 and H460. The optimized gene transfection efficacy of L3 was nearly 5.5 times more efficient than that of the popular commercially available gene delivery agent Lipofectamine 2000™ in human lung carcinoma cells A549. In addition, since L1 and L3 had nearly no gene transfection performance in normal cells HEK293, these cationic lipids showed tumor cell-targeting property to a certain extent. No significant cytotoxicity was found for the lipoplexes formed by L1–L3, and their cytotoxicities were similar to or slightly lower than the lipoplexes prepared from Lipofectamine 2000™.ConclusionNovel cyclen-based cationic lipids for effective in vitro gene transfection were founded, and these studies here may extend the application areas of macrocyclic polyamines, especially for cyclen.
Highlights
Gene therapy, as a promising therapeutics to treat genetic or acquired diseases, has received significant attentions in the past several decades due to its advantages over traditional therapies [1,2,3]
Cyclen derivatives, which was extensively studied in our group for their synthesis and interaction with DNA [21,28,29,30,31] was introduced here because protonated cyclen has pH-dependent charged nitrogen, centralized amine density and non-folded conformations which would make against their interactions with DNA [18]
The concentration of DNA was 0.5 mg/well and the cytotoxicity of lipoplexes prepared from Lipofectamine 2000TM was used as control (Lipo)
Summary
As a promising therapeutics to treat genetic or acquired diseases, has received significant attentions in the past several decades due to its advantages over traditional therapies [1,2,3]. The visible fruits of such intense global efforts toward developing safe and efficient cationic lipids for use in gene delivery are a number of commercially available cationic lipid-based transfection kits such as Lipofectamine and Lipofectamine 2000TM [9]. Those cationic lipids were still far from the requirement of gene therapy because of their relative low transfection efficiency and potential cytotoxicity [10,11]. Delivery problems limited their clinical application, and new gene delivery vehicles with low cytotoxicity and high transfection efficiency are greatly required
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