論文 - 詳細
| RRC ID | 65039 |
|---|---|
| 著者 | Hagiwara K, Kishimoto S, Ishihara M, Koyama Y, Mazda O, Sato T. |
| タイトル | In vivo gene transfer using pDNA/chitosan/chondroitin sulfate ternary complexes: influence of chondroitin sulfate on the stability of freeze-dried complexes and transgene expression in vivo. |
| ジャーナル | J Gene Med |
| Abstract |
BACKGROUND:Chitosan has been investigated as a promising nonviral vector. However, several problems still remain, such as a relatively low transfection efficiency and instability under physiological conditions. We previously demonstrated that a chondroitin sulfate (CS) coating enhanced the transfection efficiency and physicochemical stability of plasmid DNA (pDNA)/chitosan complexes in vitro. In the present study, the effects of coating pDNA/chitosan complexes with CS on the stability in freeze-dry rehydration processes and gene expression in vivo were investigated. METHODS:Freeze-drying storage at -20 °C, 4 °C, or room temperature, freezing storage at -20 °C, or liquid storage at 4 °C or room temperature, were examined for preservation conditions of pDNA/chitosan/CS ternary complexes by a gel retardation assay, measurements of sizes and zeta potentials, and a luciferase assay. Moreover, to determine the transfection efficiency of the ternary complexes in vivo, suicide gene therapy was carried out in Huh-7-implanted mice using herpes simplex virus thymidine kinase coding pDNA and ganciclovir. RESULTS:The freeze-dried pDNA/chitosan/CS ternary complexes showed sufficient cell transfection ability in vitro and in vivo. In addition, ternary complexes were associated with a significant suppression of tumor growth and a histopathologically high anti-tumor effect by intratumoral injection to tumor-bearing mice. CONCLUSIONS:The CS coating enhanced the preservation stability of the pDNA/chitosan complexes after freeze-drying-rehydration and their transgene expression in vivo. |
| 巻・号 | 15(2) |
| ページ | 83-92 |
| 公開日 | 2013-2-1 |
| DOI | 10.1002/jgm.2694 |
| PMID | 23307647 |
| MeSH | Animals Antineoplastic Agents / pharmacology Cell Line, Tumor Cell Survival / drug effects Chitosan / chemistry* Chondroitin Sulfates / chemistry* DNA / chemistry* DNA / genetics Electrophoretic Mobility Shift Assay Freeze Drying Ganciclovir / chemistry Gene Expression Gene Transfer Techniques* Genes, Transgenic, Suicide* Genetic Therapy / methods Growth Inhibitors / pharmacology Humans Injections, Intralesional Luciferases / analysis Male Mice Particle Size Plasmids / chemistry* Simplexvirus / enzymology Simplexvirus / genetics Thymidine Kinase / genetics Transfection beta-Galactosidase / analysis beta-Galactosidase / genetics |
| IF | 3.258 |
| リソース情報 | |
| ヒト・動物細胞 | HuH-7(RCB1366) |