鱼精蛋白-siRNA不同形貌复合体的制备, 结构及药效学分析

Protamine-siRNA complexes of different morphologies were prepared by vortex mixing different mass ratios of protamine and siRNA at room temperature, which could be resolved by polyacrylamide gel electrophoresis with silver staining. The protamine-siRNA complexes were observed under atomic force and transmission electron microscopy, respectively. It was found that similarly shaped protamine-siRNA complexes could be formed by protamine with different siRNA molecules, indicated that the interaction between protamine and siRNA was independent on the base composition of siRNA. Moreover, the interactions between the protamine and the siRNA molecules were characterized by ionic probes, including Mg²⁺ etc. And the morphologies, the particle sizes as well as the structural properties of the protamine-siRNA complexes were further characterized using atomic force microscope, dynamic light scattering and transmission electron microscope, respectively. The results show that protamine formed toroid complexes and fibrous composites with siRNA molecules respectively in the light of the different mass ratios between protamine and siRNA. In the meantime, the phagocytosis and the pharmacodynamics of protamine-siRNA complexes of different morphologies were compared and measured using melanoma cells.