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目的:通过自组装方法使用蛋白质分子修饰聚酰胺-胺树状大分子(PAMAM)与DNA形成的转染复合物,并考察其性质。方法:使用人血白蛋白(HSA)和表皮生长因子(EGF)修饰PAMAM-DNA转染复合物,测定复合物粒径及Zeta电位;通过DNA固缩程度测定试验和DNaseI消化试验考察修饰后复合物的稳定性;HEK 293T细胞与MCF-7细胞转染质粒,测定其荧光素酶表达水平;MTT检测修饰后复合物对细胞毒性的变化。结果:自组装修饰后的复合物的粒径无显著变化,Zeta电位显著降低,性质稳定;HSA修饰可显著提高复合物在HEK 293T和MCF-7细胞中的转染效率,EGF修饰仅显著提高复合物在MCF-7细胞中的转染效率;两种修饰都能降低PAMAM-DNA复合物对细胞的毒性。结论:自组装修饰方法快速、简便、有效,不影响PAMAM-DNA复合物的稳定性,并且能够实现提高转染效率、靶向递送、降低毒性等目的。
OBJECTIVE: To characterize the transfection complex of polyamide-amine dendrimer (PAMAM) and DNA by protein assembly using self-assembly method. Methods: The PAMAM-DNA transfection complex was modified with human serum albumin (HSA) and epidermal growth factor (EGF), and the particle size and Zeta potential of the complex were measured. The degree of DNA zeta potential HEK 293T cells and MCF-7 cells were transfected with plasmids to determine the luciferase expression level. MTT was used to detect the cytotoxicity of the modified complexes. Results: There was no significant change in the size of the composite after self-assembly, and the Zeta potential was significantly decreased. The HSA modified transfection efficiency of HEK 293T and MCF-7 cells was significantly increased The transfection efficiency of the complex in MCF-7 cells; both modifications reduce the cytotoxicity of the PAMAM-DNA complex. Conclusion: The self-assembly modification method is rapid, simple and effective, does not affect the stability of PAMAM-DNA complex, and can achieve the purpose of improving transfection efficiency, targeted delivery, reduce toxicity and so on.