Suppression of VEGF transcription in renal cell carcinoma cells using pyrrole-imidazole hairpin polyamides targeting the hypoxia responsive element

使用靶向缺氧反应元件的吡咯-咪唑发夹聚酰胺抑制肾细胞癌细胞中的 VEGF 转录

• 批准号: 14370505
• 负责人: KAGEYAMA Yukio
• 金额: $ 3.78万
• 依托单位: Graduate School Tokyo Medical and Dental 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 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370505/
• 关键词: Polyamide; Renal cell cancer; Hypoxia inducible factor; VEGF; Neovascularization; 低酸素; 転写因子; 血管内皮成長因子; 低酸素応答転写因子; HRE; 血管新生抑制; ピロールイミダゾールポリアミド; 腎癌

Hypoxia inducible factor (HIF), a master regulator of critical genes for cell survival under hypoxia, is known to be related to tumorigenesis and progression of renal cell carcinoma. N-methylpyrrole (Py)-N-methylimidazole (Im) hairpin polyamides are synthetic organic compounds which recognize and bd to minor groove of specific DNA sequences. We synthesized three Py-Im hairpin polyamides targeting the flanking sequences of hypoxia responsive element (a binding site of HIF) in the promoter region of the vascular endothelial growth factor (VEGF) gene. Effects of the polyamides on HIF-induced transcription was evaluated by luciferase assay using a reporter plasmid containing VEGF promoter. Real time reverse transcriptase polymerase chain reaction and enzyme-linked immunosolvent assay were performed to examine effects of the polyamides on transcription and secretion of VEGF in A498 renal cell carcinoma cells which has a frame shift mutation in the von Hippel-Lindau gene. Combinations of three Py-Im hairpin polyamides suppressed HIF-induced transcription in reporter assays using 293 cells, and successfully suppressed transcription and translation of the VEGF gene in A498 cells. The approach using Py-Im hairpin polyamides may provide a new strategy for the treatment of renal cell carcinoma that is resistant to conventional chemotherapy or irradiation.

缺氧诱导因子（HIF）是缺氧条件下细胞生存关键基因的主要调节因子，已知与肾细胞癌的肿瘤发生和进展有关。 N-甲基吡咯 (Py)-N-甲基咪唑 (Im) 发夹聚酰胺是合成有机化合物，可识别并结合特定 DNA 序列的小沟。我们合成了三种 Py-Im 发夹聚酰胺，靶向血管内皮生长因子 (VEGF) 基因启动子区域缺氧反应元件（HIF 的结合位点）的侧翼序列。使用含有 VEGF 启动子的报告质粒通过荧光素酶测定评估聚酰胺对 HIF 诱导转录的影响。采用实时逆转录酶聚合酶链反应和酶联免疫溶剂测定法检测聚酰胺对 von Hippel-Lindau 基因移码突变的 A498 肾细胞癌细胞中 VEGF 转录和分泌的影响。在使用 293 细胞的报告基因测定中，三种 Py-Im 发夹聚酰胺的组合抑制了 HIF 诱导的转录，并成功抑制了 A498 细胞中 VEGF 基因的转录和翻译。使用 Py-Im 发夹聚酰胺的方法可能为治疗对传统化疗或放疗耐药的肾细胞癌提供新策略。

项目成果

Koga F, Kawakami S, Fujii Y, Saito K, Iwai A, Kumagai J, Takizawa T, * Ando N, Kageyama Y, Kihara K: "Impaired p63 expression associates with poor^〓 prognosis and uroplakin III expression in invasive urothelial carcinoma of^〓 the bladder."Clin Cancer Res.

Koga F、Kawakami S、Fujii Y、Saito K、Iwai A、Kumagai J、Takizawa T、* Ando N、Kageyama Y、Kihara K：“p63 表达受损与侵袭性尿路上皮癌的不良^〓预后和尿斑蛋白 III 表达相关^〓膀胱。”临床癌症研究。

Prognostic significance of endothelial per-arnt-sim domain protein 1/hypoxia-inducible factor-2a expression in a subset of tumor associated macrophages in invasive bladder cancer.

浸润性膀胱癌肿瘤相关巨噬细胞子集中内皮 per-arnt-sim 结构域蛋白 1/缺氧诱导因子 2a 表达的预后意义。

• 发表时间: 2004
• 期刊: J Urol 171
• 作者: Koga F;Kageyama Y;Kawakami S;Fujii Y;Hyochi N;Ando N;Takizawa T;Saito K;Iwai A;Masuda H;Kihara K.
• 通讯作者: Kihara K.

腎尿路腫瘍と低酸素応答転写因子

肾泌尿道肿瘤和缺氧反应转录因子

• 发表时间: 2002
• 期刊: 泌尿器外科 15
• 作者: Fukata S;Inoue K;Kamada M;Kawada C;Furihata M;Ohtsuki Y;Shuin T.;影山幸雄
• 通讯作者: 影山幸雄

Saito K, Fujii Y, Kawasaki S, Hayashi T, Arisawa C, Koga F, Kageyam Y, Kihara: "Increased expression of sialyl-Lewis A correlates with poor survival in upper urinary tract urothelial cancer patients]."Anticancer Res. 23. 3441-3446 (2003)

Saito K、Fujii Y、Kawasaki S、Hayashi T、Arisawa C、Koga F、Kageyam Y、Kihara：“唾液酸-Lewis A 表达增加与上尿路尿路上皮癌患者生存率低相关]。”Anticancer Res。

Prognostic significance of endothelial Per-Arnt-sim domain protein 1/hypoxia-inducible factor-2alpha expression in a subset of tumor associated macrophages in invasive bladder cancer.

浸润性膀胱癌肿瘤相关巨噬细胞子集中内皮 Per-Arnt-sim 结构域蛋白 1/缺氧诱导因子 2α 表达的预后意义。

• 发表时间: 2004
• 期刊: J Urol 171
• 作者: Koga F;Kageyama Y et al.
• 通讯作者: Kageyama Y et al.