Molecular constructions of oligodeoxynucleotides binding to bFGF targeting for angiogenesis

血管生成受肝素结合生长因子如碱性成纤维细胞生长因子的调控。在本研究中，我们研究了硫代寡脱氧核苷酸对碱性成纤维细胞生长因子诱导的血管生成的体外和体内影响。反义硫代寡核苷酸被广泛用作基因表达的特异性抑制剂。因为硫代磷酸酯型寡核苷酸是多阴离子，它们也可以结合许多肝素结合蛋白。在电泳迁移率转移试验中，硫代寡核苷酸以剂量依赖但序列独立的方式结合碱性成纤维细胞生长因子。然而，双链磷硫酸寡核苷酸dna不影响复合体的形成。在基质基质上，我们观察到含有10 μ m碱性成纤维细胞生长因子、血管内皮生长因子或核因子kappa B反义以及义磷酸的人脐带内皮细胞形成管的比例不到50%，而磷酸二酯型寡脱氧核苷酸不受影响。添加1 ug/ml碱性成纤维细胞生长因子可以克服这种影响。硫代磷酸酯寡脱氧核苷酸对碱性成纤维细胞生长因子诱导的兔角膜新生血管也有抑制作用。碱性成纤维细胞生长因子与含有4个连续鸟苷残基的核因子- κ B反义序列孵育后，可明显消除碱性成纤维细胞生长因子的作用。在白化病大鼠中，核因子- κ B反义磷酸寡脱氧核苷酸在碱性成纤维细胞生长因子依赖的光感受器中也表现出较低的拯救评分，因为它在持续的光暴露下被降解。然而，对碱性成纤维细胞生长因子有活性的反义硫代寡脱氧核苷酸比含有4个鸟苷的硫代寡脱氧核苷酸对血管生成的抑制作用更强。由于碱性成纤维细胞生长因子在血管生成中的重要作用，一些硫代磷寡脱氧核苷酸可能通过多阴离子硫代磷效应和序列特异性反义机制的结合而成为有效的抗血管生成化合物。少

Angiogenesis is regulated by heparin-binding growth factors such as basic fibroblast growth factor. In this study, we investigated the effects of phosphorothioate oligodeoxynucleotides on basic fibroblast growth factor induced angiogenesis in vitro and in vivo. Antisense phosphorothioate oligonucleotides are widely used as specific inhibitors of gene expression. Because phosphorothioate-type oligonucleotides are polyanions, they can also bind many heparin-binding proteins. In an electrophoretic mobility shift assay, phosphorothioate oligonucleotides bound basic fibroblast growth factor in a dose-dependent but sequence-independent manner. However, double-strand phosphothioate oligonucleotide-DNAs did not affect complex formation. On a basement matrix using a Matrigel matrix, we observed less than 50% tube formation by human umbilical endothelial cells with 10 uM basic fibroblast growth factor, vascular endothelial growth factor or Nuclear factor kappa B antisense as well as sense phosph … More orotiate oligodeoxynucleotides, while phosphodiester-type oligodeoxynucleotides were not affected. This effect could be overcome by addition of 1 ug/ml basic fibroblast growth factor. The phosphorothioate but not phosphodiester oligodeoxynucleotides also inhibited the basic fibroblast growth factor-induced rabbit corneal neovascularization. The incubation of basic fibroblast growth factor with nuclear factor-kappa B antisense sequence containing four contiguous guanosine residues significantly abrogated the effects of basic fibroblast growth factor. The nuclear factor-kappa B antisense phosphorotioate oligodeoxynucleotides also showed a low rescue score for basic fibroblast growth factor-dependent photoreceptor rescue due to its degradation by constant light exposer in albino rats. However, antisense phosphorothioate oligodeoxynucleotides active against basic fibroblast growth factor inhibited angiogenesis more strongly than the phosphorothioats oligodeoxynucleotides containing four guanosines. Because of the important role basic fibroblast growth factor plays in angiogenesis, some phosphorothioate oligodeoxynucleotides may serve as potent anti-angiogenic compounds acting through a combination of polyanionic phosphorothioate effects and a sequence-specific antisense mechanism. Less