聚乙烯亚胺/环糊精聚合物偶合雷公藤内酯醇的体外抗肿瘤活性研究

Anticancer effect of triptolide-polyethylenimine-cyclodextrin in vitro

目的:合成并表征雷公藤内酯醇-聚乙烯亚胺-环糊精聚阳离子复合物,考察雷公藤内酯醇-聚乙烯亚胺-环糊精在几种不同癌细胞上的抗癌活性和siRNA携带能力。方法:通过N,N'-羰基二咪唑将雷公藤内酯醇偶合到聚乙烯亚胺-环糊精载体材料上,制成雷公藤内酯醇-聚乙烯亚胺-环糊精纳米复合物。用1H-NMR、FT-IR和XRD对该材料进行了谱学表征,用MTT、划痕实验和伊红-苏木精染色实验对其细胞毒性和体外抗肿瘤活性进行了评估,并用凝胶电泳实验、粒径和表面电势的测定及携带siRNA的细胞荧光染色实验,初步研究了聚合物的siRNA压缩能力与携带能力。结果:合成了雷公藤内酯醇-聚乙烯亚胺-环糊精聚阳离子复合物,且雷公藤内酯的接入率为10%(w/w)。聚合物在N/P为5时具有较好的siRNA压缩能力,形成粒径约为(300±15)nm和表面电荷为(8±2.5)mV的微粒,细胞毒性实验、划痕实验及HE染色实验表明雷公藤内酯醇-聚乙烯亚胺-环糊精聚合物毒性降低,具有较好的抑制细胞转移能力,能有效携带siRNA并进入细胞中。结论:雷公藤内酯醇-聚乙烯亚胺-环糊精可在体外抑制癌细胞的生长和迁移,并可携带siRNA进入细胞内部,有望用于肿瘤的基因/药物协同治疗。

Objective： To develop a drug delivery system triptolide-polyethylenimine-cyclodextrin and to evaluate its anticancer activity in vitro.Methods： Triptolide was conjugated to polyethylenimine-cyclodextrin by N,N ′-carbonyldiimidazole to form triptolide-polyethylenimine-cyclodextrin.1H-NMR,FT-IR and XRD were used to confirm its structure.The anticancer effect of the polymer was assessed by MTT assay,erasion trace test and hematoxylin-eosin staining.The potential to condense siRNA and to delivery siRNA into cytoplasm was demonstrated by gel retardation assay,zeta-potential determination and fluorescence staining.Results： Triptolide was successfully conjugated to polyethylenimine-cyclodextrin and the conjugation rate of triptolide was 10%（w/w）.siRNA was effectively condensed by the polymer at the N/P ratio of 5,and its particle size was 300±15 nm and zeta potential was 8±2.5 mV.MTT assay,erasion trace test and hematoxylin-eosin staining revealed that triptolide-polyethylenimine-cyclodextrin had anticancer effect and low cytotoxicity to normal cells.The polymer was able to deliver siRNA to the cytoplasm effectively as demonstrated by fluorescence staining.Conclusions： Triptolide-polyethylenimine-cyclodextrin is able to inhibit the growth and migration of cancer cells in vitro and to carry siRNA into cells effectively.It is potential to be used as a novel prodrug for co-delivery of gene and drug in cancer treatment.