ESTBLISHMENT AND APPLIATCE OF NEWLY GENE THERAPY FOR INHIBITION OF TUMOR ANGIOGENESIS

抑制肿瘤血管生成新基因疗法的建立及应用

• 批准号: 12671830
• 负责人: ISHIBASHI Hiroaki
• 金额: $ 2.18万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671830/
• 关键词: ANGIOGENESIS; NEOPLASMS; GENETHERAPY

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 α (TNFα) via binding of transcription factor Sp1 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 "decoy" could block the binding of Sp1 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 Sp1 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 Sp1 could be simultaneously suppressed.

周围组织的正常微血管形成新的毛细血管网络被认为对实体瘤的生长起关键作用。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）是最有效的血管生成因子之一，是内皮细胞特异性的有丝分裂原。先前的一些研究表明，在各种体内和体外模型中，使用反义寡脱氧核苷酸或中和抗体抑制VEGF可抑制肿瘤生长。众所周知，VEGF是缺氧诱导的，最近有报道称，它是通过肿瘤坏死因子α (TNFα)的刺激，通过转录因子Sp1与VEGF启动子的结合而合成的。我们假设将合成的双链DNA（包括Spl结合位点的一致序列）作为顺式反式元件“诱饵”转染到肿瘤细胞核中可以阻断Sp1与VEGF启动子基因的结合。转染野生型Spl诱饵，而非突变型诱饵，在TNFα刺激下对培养的人癌细胞VEGF合成有明显的抑制作用。将Sp1诱骗物引入肿瘤细胞可能是一种新的有用的治疗工具，通过其对VEGF合成的抑制作用诱导肿瘤优势。此外，Spl诱饵的转移比反义寡核苷酸的转移更有效地调节肿瘤的生长和侵袭，因为Sp1介导的血管生成因子和生长及侵袭相关因子的表达并不能同时被抑制。

项目成果

T Shiratuchi et al.: "Inhibition of epidermal growth factor-induced invasion by dexamethasone in human squamous cell carcinoma cell lines"J Cell Physiol. (In press).

T Shiratuchi 等人：“人鳞状细胞癌细胞系中地塞米松抑制表皮生长因子诱导的侵袭”J Cell Physiol。

Nakagawa K, Chen Y, Ishibashi H, et al.: "Angiogenesis and its regulation : roles of vascular endothelial cell growth factor (VEGF)"Semin Thromb Hemost. 143. 201-206 (2000)

Nakakawa K、Chen Y、Ishibashi H 等人：“血管生成及其调节：血管内皮细胞生长因子 (VEGF) 的作用”Semin Thromb Hemost。

Ishibashi H, et al.: "Hypoxia-Induced Angiogenesis of Cultured Human Salivary Gland Carcinoma CellsInvolves Enhancement VEGF Production and bFGF Release"Oral Oncology. 37. 77-83 (2001)

Ishibashi H 等人：“培养人唾液腺癌细胞的缺氧诱导血管生成涉及增强 VEGF 产生和 bFGF 释放”口腔肿瘤学。

Nakagawa K, Chen Y, Ishibashi H, et al.: "Angiogenesis and its regulation : roles of vascular endothelial cell growth factor (VEGE)"Semin Thromb Hemost. 143. 201-206 (2000)

Nakakawa K、Chen Y、Ishibashi H 等人：“血管生成及其调节：血管内皮细胞生长因子 (VEGE) 的作用”Semin Thromb Hemost。

H.Ishibashi, et al: "Sp1 Decoy Transfected Cells Suppresses the Expression of VEGF, TGFβ1, and Tissue Facotr and also Cell Grwoth and Invasion Activites."Cancer Research. 60. 6531-6536 (2000)

H. Ishibashi 等人：“Sp1 诱饵转染细胞抑制 VEGF、TGFβ1 和组织因子的表达以及细胞生长和侵袭活动。”癌症研究 60. 6531-6536 (2000)。