Development of Gene Therapy with Nanotechnology derived Intelligent Drug Deliver Systems

我们开发了多离子复合物与DNA和多阳离子基因传递系统。选择聚乙烯亚胺进行多阳离子化，对转染癌细胞具有质子反应作用。聚乙烯-乙二醇结合羧基链也被用于包覆带阳离子的DNA/多阳离子配合物。为了提高转染效率，我们还检测了与流感病毒分析构建的pH敏感肽JTS-1。聚乙烯亚胺与DNA复合物的荧光素酶活性为1 × 10^9 IU/mg。用PEG-C包覆DNA/多离子复合物时，荧光素酶活性提高到3 × 10^9 IU/mg，这是由于PEG-C包覆后复合物的压实性和稳定性增强，从而提高了转染效率。将JTS-1加入到DNA/聚乙烯亚胺/乳糖结合的PEG-C复合物中，很容易逃脱溶酶体的裂解并将DNA转染到细胞质中，获得6 × 10^9 IU/mg的荧光素酶活性。肿瘤内注射β-gal DNA质粒/聚乙烯亚胺/乳糖结合PEG-C/JTS-1复合物对胃癌荷瘤小鼠模型转染β-gal基因的影响对奥沙利铂或顺铂敏感或耐药的癌细胞进行DNA微阵列分析发现，在奥沙利铂或顺铂耐药的癌细胞上，Tubrin特异性Chaperon E基因和CBP/ p300相互作用反激活子（CITED2）基因增加。

We develop polyion complex with DNA and polycation for gene delivery systems. Polyethyleneimine was selected for polycation that have protonsponde effect on transfection to cancer cells. Polyethylene-glycol binding carboxyl chain was also used for coating the cationic charged DNA/polycation complex. To increase the transfection efficacy, we also examined pH sensitive peptide, JTS-1 that constructed with analysis of Influenza virus. Polyethyleneimine and DNA complex showed 1 × 10^9 IU/mg luciferase activity. In case of coating the DNA/polyion complex with PEG-C, the luciferase activity was increase to 3 × 10^9 IU/mg, because of the compaction and incresent of stability with addition of PEG-C to complex, so the transfection efficacy was increased. When addition of JTS-1 to the DNA/polyethyleneimine/lactose-binding PEG-C complex, it is easily to escape the lysis of lysosome and transfect the DNA to cytoplasm, and 6 × 10^9 IU/mg luciferase activity was obtained. Increasing of transfection efficacy of β-gal gene by intratumor injection of β-gal DNA plasmid/Polyethyleneimine/lactose-binding PEG-C/JTS-1 complex on gastric cancer tumor bearing mice model.According to the analysis of DNA microarray with ancer cell lines that were sensitive or resistant to Oxaliplatin or Cisplatin, Tubrin specific Chaperon E gene and CBP/P300-interacting transactivator (CITED2) gene were increased on Oxaliplatin or Cisplatin resistant cancer cells.