Newly Gene Therapy Targetting Inhibition of the Tumor Angiogenesis Using Decoy System

利用诱饵系统抑制肿瘤血管生成的新基因疗法

• 批准号: 14370605
• 负责人: ISHIBASHI Hiroaki
• 金额: $ 9.02万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370605/
• 关键词: NEOPLASMS; ANGIONESIS; GENE THERAPY; 腫瘍血管新生

The development of newly capillary networks from the normal microvasculature of the surrounding tissue has thought to play a. critical role For the growth of the solid tumors. Tumor cells influence the angiogenesis by stimulation of endothelial cells with producing several angiogenic factors. Among them, vascular endothelial growth factor(VEGF) is one of the most potent angiogenic factors, and endothelial cell specific mitogen. Several previous studies suggested that inhibition of VEGF using antisense oligodeoxynucleotides or neutralizing antibodies against VEGF suppressed tumor growth in various in vivo and in vitro models. VEGF is well known to be hypoxia-inducible. and has been recently reported to be synthesized by stimulation with tumor necrosis factor a(TNFα) via binding of transcription factor Spl to the VEGF promoter. We hypothesized that transfection into the tumor cells nucleus of the synthetic double stranded DNA including consensus sequence of binding site of the Spl as cis-trans element メdecoyモ could block the binding of Spl to the VEGF promoter gene. Transfection of wild type Spl decoy, but mutant type decoy, revealed prominent inhibitory effects of VEGF synthesis of cultured human carcinoma cells stimulated by TNFα. This Spl decoy introduction into tumor cells may be a novel and useful therapeutic tool for induction of tumor domancy by its inhibitory effect on VEGF synthesis. In addition, the transfer of the Spl decoy would be more effective for regulating tumor growth and invasion than that of antisense oligonucleotide. since not all angiogenic factors but also growth and invasion related factors expression modulated by Spl could be simultaneously suppressed.

从周围组织的正常微血管系统发展出新的毛细血管网络被认为是一种重要的机制。对实体瘤的生长起着关键作用。肿瘤细胞通过刺激内皮细胞产生多种血管生成因子来影响血管生成。其中，血管内皮生长因子（VEGF）是最强的血管生成因子之一，也是内皮细胞特异性的有丝分裂原。几个先前的研究表明，使用反义寡脱氧核苷酸或针对VEGF的中和抗体抑制VEGF在各种体内和体外模型中抑制肿瘤生长。众所周知，VEGF是缺氧诱导的。并且最近已经报道通过用肿瘤坏死因子α（TNFα）刺激经由转录因子Spl与VEGF启动子的结合来合成。我们假设，将合成的包含Spl结合位点共有序列的双链DNA作为顺反元件的诱饵质粒转染到肿瘤细胞核中，可以阻断Spl与VEGF启动子基因的结合。转染野生型Spl诱饵，但突变型诱饵，显示显着抑制TNFα刺激的培养的人癌细胞的VEGF合成的效果。这种Spl诱饵引入肿瘤细胞可能是一种新的和有用的治疗工具，用于通过其对VEGF合成的抑制作用诱导肿瘤优势。此外，Spl诱骗物的转移将比反义寡核苷酸更有效地调节肿瘤的生长和侵袭。因为并非所有血管生成因子以及受Spl调节的生长和侵袭相关因子的表达都可以同时受到抑制。

项目成果

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Onimaru.M 等人：“成纤维细胞生长因子-2 基因转移可以刺激肝细胞生长因子的表达，无论缺血肢体中缺氧介导的下调如何”循环研究。

Shiratuchi, T., Ishibashi, H.et al.: "Inhibition of epidermal growth factor-induced invasion by dexamethasone and AP-1 decoy in human squamous cell carcinoma cell lines"J Cell Physiol. 93. 340-348 (2002)

Shiratuchi, T., Ishibashi, H.等人：“人鳞状细胞癌细胞系中地塞米松和 AP-1 诱饵对表皮生长因子诱导的侵袭的抑制”J Cell Physiol。