Gene transfer system mediated by PEI-cholesterol lipopolymer with lipid microbubbles.
- Author:
Yong-Nan JIANG
1
;
Hong-Ying MO
;
Jian-Hai CHEN
Author Information
1. Department of Pharmacy, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
- Publication Type:Journal Article
- MeSH:
1,2-Dipalmitoylphosphatidylcholine;
chemistry;
Breast Neoplasms;
pathology;
Cell Line, Tumor;
Cell Survival;
Cholesterol;
chemistry;
Contrast Media;
DNA;
chemistry;
genetics;
Female;
Fluorocarbons;
chemistry;
Gene Transfer Techniques;
Genetic Vectors;
Green Fluorescent Proteins;
metabolism;
Humans;
Lipids;
chemistry;
Lung Neoplasms;
pathology;
Microbubbles;
Phosphatidylethanolamines;
chemistry;
Plasmids;
Polyethylene Glycols;
chemistry;
Polyethyleneimine;
chemistry;
Transfection;
methods
- From:
Acta Pharmaceutica Sinica
2010;45(5):659-666
- CountryChina
- Language:Chinese
-
Abstract:
The properties of polyethyleneimine-cholesterol cationic lipopolymer (PEI-Chol) as gene carries and its gene transfer efficiency in vitro with lipid microbubbles were presented in this paper. PEI-Chol lipopolymer was synthesized by linking cholesterol chloroformate to the amino groups of branched poly(ethyleneimine) (PEI) of 1 800. The structure and molecular weight of PEI-Chol were confirmed by IR, 1H NMR and MADI-TOF-MS (matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry), respectively. The average molecular weight of PEI-Chol was approximately 2 000. The gene delivery system of bubble/PEI-Chol/DNA was constructed by mixed PEI-Chol/pDNA (N/P 10:1) complexes with lipid microbubbles (2-8 microm) which were prepared by DPPC, DSPE-PEG2000 and perfluoropropane with the reverse phase evaporation technique. pEGFP-Cl (enhanced green fluorescent protein) was used as report gene to investigate the DNA condensing ability of PEI-Chol lipopolymer by agarose gel electrophoresis. And their cytotoxicity and in vitro transfer efficiency of different complexes were compared with each other in A549 and MCF-7. The results indicated PEI-Chol lipopolymer can condense plasmid DNA when N/P ratio upto 4, PEI-Chol complexes and bubble/PEI-Chol/DNA complexes were nontoxic to A549 and MCF-7 when formulated at the N/P ratio of 10/1 as determined by MTT assay. This bubble/PEI-Chol/DNA delivery system provided good transfer efficiency with other desirable characteristics such as against-precipitation of plasma proteins. In conclusion, bubble/PEI-Chol/DNA complex is a novel non-viral gene delivery system.
