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目的 :利用肝细胞膜存在的去唾液酸糖蛋白受体 ,合成一种能够携带目的DNA分子特异性富集到肝组织中的载体。方法 :在弱碱性溶液中 ,在氰基硼氢化钠作用下 ,通过还原氨化法进行乳糖 (Lac)与聚赖氨酸 (PLL)的共价连接 ,SephadexG 10分离纯化产物 ;采用真核细胞表达基因 β 半乳糖苷酶报道基因 ,在细胞培养中和小鼠体内观察乳糖 聚赖氨酸复合物 (Lac PLL)对肝细胞的导向能力。结果 :Lac在氰基硼氢化钠作用下能有效地结合到PLL分子上 ,Lac与PLL的摩尔比为 16∶1;Lac PLL复合物在体外能够与DNA分子形成稳定偶联物 ,使其所偶联的DNA分子有效地进入到肝癌细胞 ,并使相应基因在细胞中表达。小鼠尾静脉注射DNA/Lac PLL偶联物10 μg后 ,DNA能有效地在肝组织中富集并在肝细胞中表达。结论 :Lac PLL是一种有效的对目的DNA分子具有肝细胞特异性导向能力的载体。
OBJECTIVE: To synthesize a carrier capable of carrying specific target DNA molecules to liver tissue by using the asialoglycoprotein receptor present in the liver cell membrane. METHODS: Covalent attachment of lactose (Lac) to polylysine (PLL) was achieved by reductive amination under the action of sodium cyanoborohydride in weak alkaline solution. Sephadex G 10 was used to separate the purified product. The eukaryotic The cells express gene β-galactosidase reporter gene and observe the ability of Lac PLL to guide hepatocytes in cell culture and in mice. Results: Lac could effectively bind to the PLL molecule under the action of sodium cyanoborohydride. The molar ratio of Lac to PLL was 16:1. Lac PLL complex could form a stable conjugate with DNA molecule in vitro, Coupled DNA molecules efficiently enter liver cancer cells and express the corresponding genes in the cells. After the tail vein of mice was injected with 10 μg DNA / Lac PLL conjugate, the DNA was efficiently enriched in liver tissue and expressed in hepatocytes. CONCLUSION: Lac PLL is a potent carrier of hepatocyte-specific targeting to DNA molecules of interest.