Molecular constructions of oligodeoxynucleotides binding to bFGF targeting for angiogenesis

结合 bFGF 靶向血管生成的寡脱氧核苷酸的分子结构

基本信息

批准号：
09470172
负责人：
KITAJIMA Isao
金额：
$ 7.68万
依托单位：
Kagoshima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

项目摘要

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

血管生成受肝素结合生长因子（例如碱性成纤维细胞生长因子）的调节。在本研究中，我们研究了硫代磷酸寡脱氧核苷酸对体外和体内碱性成纤维细胞生长因子诱导的血管生成的影响。由于硫代磷酸酯型寡核苷酸是聚阴离子，因此它们还可以结合许多肝素结合蛋白。电泳迁移率变动测定中，硫代磷酸寡核苷酸以剂量依赖性但与序列无关的方式结合碱性成纤维细胞生长因子，然而，在使用基质胶基质的基底基质上，双链硫代磷酸寡核苷酸-DNA 不影响复合物的形成。含有 10 uM 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 的人脐带内皮细胞的管形成率超过 50%反义和正义磷酸寡脱氧核苷酸，而磷酸二酯型寡脱氧核苷酸不受影响，可以通过添加 1 ug/ml 碱性成纤维细胞生长因子来克服，硫代磷酸酯（而非磷酸二酯寡脱氧核苷酸）也抑制碱性成纤维细胞生长因子。 -诱导兔角膜新生血管形成。碱性成纤维细胞生长因子与核因子-κB的孵育。含有四个连续鸟苷残基的反义序列显着消除了碱性成纤维细胞生长因子的作用，核因子-κB反义硫代磷酸寡脱氧核苷酸由于其在恒定光照下被降解，因此对于碱性成纤维细胞生长因子依赖性光感受器的拯救也表现出较低的拯救分数。然而，反义硫代磷酸寡脱氧核苷酸对碱性成纤维细胞生长因子具有抑制作用。由于碱性成纤维细胞生长因子在血管生成中发挥重要作用，一些硫代磷酸寡脱氧核苷酸可以通过聚阴离子硫代磷酸酯效应和序列特异性反义机制的组合发挥作用，作为有效的抗血管生成化合物。。较少的