Study on the preparation of a novel nonviral polycation nanostructured lipid carriers for gene delivery and its possible intracellular mechanism

摘要

A novel nonviral gene delivery system was developed containing cetyl PEI, triolein and dioleoyl phosphatidylethanolalmine (DOPE) prepared by solvent emulsifying evaporation method into the polycation nanostructured lipid carrier system (PNLC). The PNLC system was characterized and tranfection of green fluorescent protein plasmid (pEGFP-N2) and luciferase plasmid (pGL-luc) was carried out in comparison with the commercial LipofectamineTM 2000 performed as the positive control. Subsequently, the mode of internalization of PNLC/DNA complexes in SPC cells and its cytosolic transfer to the nucleus accompanied with microtubules and motor proteins were investigated as well. Results showed that the transfection efficiency of the PNLC system was higher than LipofectamineTM 2000 and triolein could heighten the transfection efficiency in SPC cells. Furthermore, the transfection efficiency of the PNLC system was enhanced in the presence of 10% serum, instead of dramatically decreased as the positivecontrol. Data showed that the clathrin-dependent pathway was the main contributor to the successful transfection mediated by PNLC system in SPC cells, and the PNLC/DNA complexes in cytoplasma were transferred in endosome undergoing motor protein driven movement guided by microtubules. Thus, PNLC represents a novel system useful for transfection and gene therapy.