RESUMO
High transfection efficiency and low cytotoxicity are the two key factors to be considered in the design of gene carriers. Herein, a novel and versatile gene carrier (PLL-RT) was prepared by introducing "molecular string" RT (i.e., p-toluylsulfonyl arginine) onto the polylysine backbone. The introduction of RT string contributed to the formation of multiple interactions between the polycationic gene carriers and cell membrane or DNA, as well as adopting α-helix conformation, all of which would be beneficial to enhance the gene transfection. In addition, RT string grafted onto other polycations such as hyperbranced PEI25k and dendrimer PAMAM could also acquire improved transfection efficiency and low cytotoxicity. Moreover, PLL-RT presented significant tumor inhibition effect in vivo. This work provided an effective strategy for constructing novel gene carriers with high transfection and low cytotoxicity.
Assuntos
DNA/genética , Técnicas de Transferência de Genes , Polilisina/análogos & derivados , Tosilarginina Metil Éster/análogos & derivados , Animais , Cardiolipinas/metabolismo , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , DNA/metabolismo , Endocitose/fisiologia , Endossomos/metabolismo , Feminino , Humanos , Membranas Artificiais , Camundongos Endogâmicos BALB C , Conformação Molecular , Neoplasias/terapia , Tamanho da Partícula , Polilisina/síntese química , Polilisina/metabolismo , Polilisina/toxicidade , RNA Interferente Pequeno/administração & dosagem , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , RNA Interferente Pequeno/uso terapêutico , Tosilarginina Metil Éster/síntese química , Tosilarginina Metil Éster/metabolismo , Tosilarginina Metil Éster/toxicidadeRESUMO
High efficiency and serum resistant capacity are important for gene carrier in vivo usage. In this study, transfection efficiency and cell toxicity of polyethylenimine (PEI) (branched, Mw = 25K) was remarkably improved, when mixed with polyanion (polyethylene glycol-polyglutamic acid (PEG-PLG) or polyglutamic acid (PLG)). Different composite orders of PEI, polyanion, and gene, for example, PEI is first complexed with DNA, and then with polyanion, or PEI is first complexed with polyanion, and then with DNA, were studied. Results showed that only the polyanion/PEI complexes exhibited additional properties, such as decreased pH, resulting in increased particle size, as well as enhanced serum resistance capability and improved tumor accumulation. The prepared gene carrier showed excellent antitumor effect, with no damage on major organs, which is suitable for in vivo gene antitumor therapy.