Hemmati Mohammad, Najafi Farhood, Shirkoohi Reza, Moghimi Hamid Reza, Zarebkohan Amir, Kazemi Bahram
a Biomedical Engineering and Medical Physics Department, Faculty of Medicine, Shahid Beheshti University of Medical Sciences , Tehran , Iran.
b Department of Resin and Additives , Institute for Color Science and Technology , Tehran , Iran.
Drug Deliv. 2016 Oct;23(8):2956-2969. doi: 10.3109/10717544.2015.1132796. Epub 2016 Aug 5.
hPAMAM/DNA polyplexes, compared to viral vectors, display unique characteristics including more safety, less immune response outcomes, a simpler synthesis and an easier process. Given the importance of these polymers, hPAMAM coated with the PEGDGA copolymer was developed as a promising non-viral gene carrier. In the present study, a new complex of hPAMAM, PEGDGA-modified hyperbranched polyamidoamine (hPAMAM), was established as a versatile non-viral gene vector. The hPAMAM polymer was synthesized by using a modified one-pot method. The resulting hPAMAM-PEGDGA polymer was able to efficiently protect encapsulated-DNA against degradation for over 2 h. In addition to low cytotoxicity, the transfection efficiency of hPAMAM-PEGDGA represented much higher (p < 0.05) than that of Lipofectamine 2000 in both MCF7 and MDA-MB231 cells (an approximately 4.5-fold increase). Cellular uptake of hPAMAM-PEGDGA in MDA-MB231 cells, 254.79 ± 2.1, was significantly higher than that in MCF7 cells, 51.61 ± 6.1 (p < 0.05). EMA-labeled DNA can be clearly observed in the tumor tissue of mice receiving hPAMAM-PEGDGA/EMA-labeled DNA. However, a significant number of fluorescent spots can be found in the tumor tissue of mice receiving hPAMAM/DNA, when compared to those treated with naked hPAMAM/DNA. It has been observed that GFP is expressed more highly in hPAMAM-PEGDGA/EMA-labeled/DNA than the one in PAMAM/DNA. The results indicated that hPAMAM-PEGDGA-mediated gene delivery to breast cancer cells is a feasible and effective strategy that may offer a new therapeutic avenue as a non-viral gene delivery carrier. Notably, According to these findings, this newly-introduced copolymer, the hPAMAM-PEGDGA complex, has proved to be a promising strategy for drug or gene delivery to tissues or cell types of interest, particularly to triple-negative breast cancer.
与病毒载体相比，人源化聚酰胺-胺树形聚合物（hPAMAM）/DNA复合物体现出独特的特性，包括更高的安全性、更低的免疫反应结果、更简单的合成方法和更便捷的流程。鉴于这些聚合物的重要性，涂覆了聚乙二醇二缩水甘油丙烯酸酯（PEGDGA）共聚物的hPAMAM被开发为一种有前景的非病毒基因载体。在本研究中，一种新型的hPAMAM复合物，即PEGDGA修饰的超支化聚酰胺-胺（hPAMAM），被确立为一种通用的非病毒基因载体。hPAMAM聚合物采用改良的一锅法合成。所得的hPAMAM-PEGDGA聚合物能够有效保护包裹的DNA在2小时以上不被降解。除了低细胞毒性外，hPAMAM-PEGDGA在MCF7和MDA-MB231细胞中的转染效率均显著高于Lipofectamine 2000（p < 0.05）（约提高4.5倍）。MDA-MB231细胞对hPAMAM-PEGDGA的细胞摄取量为254.79±2.1，显著高于MCF7细胞的51.61±6.1（p < 0.05）。在接受hPAMAM-PEGDGA/EMA标记DNA的小鼠肿瘤组织中可以清晰观察到EMA标记的DNA。然而，与接受裸hPAMAM/DNA处理的小鼠相比，在接受hPAMAM/DNA的小鼠肿瘤组织中可以发现大量荧光斑点。据观察，绿色荧光蛋白（GFP）在hPAMAM-PEGDGA/EMA标记/DNA中的表达高于在PAMAM/DNA中的表达。结果表明，hPAMAM-PEGDGA介导的基因传递至乳腺癌细胞是一种可行且有效的策略，可能作为一种非病毒基因传递载体提供新的治疗途径。值得注意的是，根据这些发现，这种新引入的共聚物，即hPAMAM-PEGDGA复合物，已被证明是一种有前景的策略，可用于将药物或基因传递至感兴趣的组织或细胞类型，特别是三阴乳腺癌。
