用于小干扰RNA递送的基于超分子主客体化学组装的聚集诱导发光囊泡材料的构建及其对肿瘤细胞毒性的研究

Establishment an aggregation-induced emission vesicle material based on supramolecular host-guest chemical assembly for siRNA delivery and study on its toxicity to tumor cells

目的:构建一种基于超分子主客体化学组装的聚集诱导发光囊泡材料(AIE-HG-Vesicle)用于递送小干扰RNA(siRNA)及实现荧光成像功能,并探究其被肿瘤细胞摄取的效果及其基于siRNA对肿瘤细胞的杀伤效果。方法:通过合成聚乙烯亚胺树型分子修饰的β-环糊精(H-β-CD-dendrimer)作为主体化合物,同时合成含有四苯乙烯AIE基团的Bola型金刚烷(G-Ada-AIE)作为客体化合物,通过超分子主客体自组装过程制备得到AIE-HG-Vesicle囊泡材料用于负载siRNA。通过透射电子显微镜及动态光散射仪测试其形貌和尺寸,通过荧光分光光度计测试其聚集诱导发光性质,通过凝胶阻滞实验测试其对siRNA的负载效果,通过激光共聚焦显微镜测试负载siRNA的AIE-HG-Vesicle囊泡材料在肿瘤细胞内的递送效果,并通过MTT实验测试负载siRNA的AIE-HG-Vesicle囊泡材料对肿瘤细胞的杀伤效果。结果:AIE-HG-Vesicle具有囊泡状形态,直径约为100 nm,壁厚约为9 nm,且表面带正电荷可有效负载siRNA。负载于AIE-HG-Vesicle的siRNA表现出良好的稳定性,且负载siRNA后的AIE-HG-Vesicle可将siRNA递送到HeLa细胞内部,并可通过聚集诱导发光现象被观测到,负载siRNA后的囊泡对HeLa细胞有明显的杀伤效果。结论:构建了AIE-HG-Vesicle可用于递送siRNA,该材料具有荧光成像和基于siRNA的肿瘤细胞毒性作用,后期有望应用于肿瘤体内治疗。

ObjectiveTo establish an aggregation-induced emission vesicle material based on supramolecular host-guest chemical assembly(AIE-HG-Vesicle)for siRNA delivery and fluorescence imaging,and to explore its uptake effect by tumor cells and siRNA-based cell killing effect.MethodsBy synthesizingβ-cyclodextrin modified with polyethyleneimine dendrimer(H-β-CD-dendrimer)as a host compound and a Bola type adamantane containing tetrastyrene AIE group(G-Ada-AIE)as a guest compound,the nanovesicle material was prepared by a supramolecular host-guest self-assembly process for loading siRNA.The morphology and size of the materials were tested by transmission electron microscopy and the dynamic light scattering method.The aggregation-induced luminescence properties of the materials were investigated by fluorescence spectrophotometry.The loading effect of the material on siRNA was investigated by gel retardation experiments.The delivery effect of siRNA-loaded AIE-HG-Vesicle vesicles in tumor cells was observed by a confocal laser scanning microscope.The killing effect of siRNA-loaded AIE-HG-Vesicle vesicles on tumor cells was tested by an MTT assay.ResultsThe prepared host-guest compounds can be assembled into vesicles with a size of about 100 nm and wall thickness of 9 nm in solution,and the positively charged vesicles on the surface can efficiently load siRNA.The siRNA-loaded AIE-HG-Vesicle vesicles can deliver siRNA into HeLa tumor cells and can be observed through aggregation-induced luminescence.The siRNA-loaded vesicles have an obvious killing effect on HeLa tumor cells.ConclusionsA vesicle material with aggregation-induced luminescence properties was prepared by a method based on supramolecular host-guest chemical assembly,which can be used to deliver siRNA.The material has fluorescence imaging and siRNA-based tumor cytotoxic effects and is expected to be applied to tumor treatment in vivo.