肝细胞靶向基因药物分子构形和表达效率的研究  

Molecular conformation and expression effciency in the hepatocyte targeting gene drug

作　　者：卢放根[1] 刘小伟[1] 欧阳春晖[1] 羊东晔[1] 吴晓英[1] 

机构地区：[1]中南大学湘雅二医院消化内科,长沙410011

出　　处：《湖南医科大学学报》2002年第1期9-12,共4页Bulletin of Hunan Medical University

基　　金：国家自然科学基金 (3 95 70 3 3 5 ) ;科技部博士创新新药基金和湖南省生命科学中心资助

摘　　要：目的 :用调整肝细胞靶向基因药物分子构形的方法 ,以期提高外源转入基因在目标细胞的表达效率。方法 :加入特定辅料分子至基因药物中 ,用电镜观察其液态分子构形。在转入体外培养的人肝癌细胞株BEL 74 0 2中后 ,用ELISA方法检测基因表达产物IFN r的浓度 ,筛选最佳表达外源基因药物分子的构形。结果 :肝细胞靶向基因药物在辅料分子不同浓度中构形发生改变 ,显现出絮状、球形、串蛛状及杆状与类染色体形的混合体。筛选出了具最佳表达的杆形和类染色体混合体分子构形。该构形的药物分子在体外培养细胞中的表达显著高于脂质体转运载体携带的同一基因 ,且不需要抑制溶酶的活性。结论 :用于基因治疗的药物分子构形对其转入靶细胞的表达效率有重要的影响。杆状和类染色体复合分子构形 。Objective To modulate the molecular conformation of the hepatocyte targeting drug to increase exogenous gene expression efficiency in the targeted cell.Methods Some specific auxiliary molecules were added to the gene drug its molecular liquid conformation was investigated under the electron microscope and they were transfected into human hepatoma cell line BEL 7402 in vitro. We detected IFN γ gene expression product by ELISA and screened out the most efficient drug molecules. Results Due to various concentrations of auxiliary molecules, the conformation of hepatocyte targeting drug changed, including floccule, globulous, string beads, bacillform and chromosome like mixture. While the most efficient expression conformation was the bacilliform and chromosome like mixture and didn't need to inhibit the lysozyme activity, the expression efficiency of such drug molecules was much higher than that of the liposome targeting vector. Conclusion The molecular drug conformation had much influence on targeted gene expression efficiency in gene therapy. Bacilliform and chromosome like mixture may be the most efficient expression conformation to construct drug molecules in ASOR targeted hepatocyte gene therapy. So we put forward a new concept 'chromosome mimic conformation'in screening out the most efficient molecular drug conformation in gene therapy.

关 键 词：基因治疗 肝细胞 肝脏 表达效率 靶向基因 药物分子构形 

分 类 号：R735.7[医药卫生—肿瘤]