Ribozyme approach to study functions of vascular endothelial growth factor isoforms on the progression of non-small cell lung cancer

核酶方法研究血管内皮生长因子亚型对非小细胞肺癌进展的作用

• 批准号: 11670227
• 负责人: NAKAMURA Masato
• 金额: $ 2.5万
• 依托单位: Tokai University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11670227/
• 关键词: ribozyme; vascular endothelial growth factor; non-small cell lung cancer; isoform; angiogenesis; cancer gene therapy; 肺癌; 遺伝子導入

The specific role of the cell-associated isoform of vascular endothelial growth factor (VEGF189) has not been clarified, while five isoforms of VEGF which have different biological activities (VEGF121, VEGF145, VEGF165, VEGF189, VEGF206) are generated by alternative splicing. We used a hammerhead-type ribozyme (V189Rz) to suppress VEGF189 mRNA.The V189Rz specifically cleaved exon 6 of VEGF189 mRNA, but showed no activity against VEGF121 or VEGF165 isoforms. The V189Rz was introduced into the human non-small cell lung cancer cell line (OZ-6/VR). The expression level of VEGF189 mRNA was decreased in the OZ-6/VR, while VEGF121 and 165 expression were preserved. The OZ-6/VR xenotransplanted into nude mice showed markedly reduced vascularisation and growth, whereas the cell line did not show any decreased growth under tissue culture conditions. The OZ-6/VR cells (1×10^5/mouse) formed no tumours, whereas the parental OZ-6 cells formed huge tumours within 8 weeks. The specific suppression of VEGF189 by the ribozyme decreased vascularity and xenotranplantability of the lung cancer cell line. The cell-associated isoform of VEGF (VEGF189) might have key roles on stromal vascularisation and growth of non-small cell lung cancer xenografts in vivo.

血管内皮生长因子（VEGF 189）的细胞相关亚型的具体作用尚未阐明，而具有不同生物学活性的VEGF的五种亚型（VEGF 121、VEGF 145、VEGF 165、VEGF 189、VEGF 206）通过选择性剪接产生。我们使用锤头型核酶（V189 Rz）抑制VEGF 189 mRNA，V189 Rz特异性切割VEGF 189 mRNA的外显子6，但对VEGF 121或VEGF 165亚型没有活性。将V189 Rz导入人非小细胞肺癌细胞系（OZ-6/VR）。在OZ-6/VR中VEGF 189 mRNA的表达水平降低，而VEGF 121和165的表达保持不变。异种移植到裸鼠中的OZ-6/VR显示出显著减少的血管形成和生长，而细胞系在组织培养条件下没有显示出任何生长减少。OZ-6/VR细胞（1×10^5/小鼠）没有形成肿瘤，而亲代OZ-6细胞在8周内形成了巨大的肿瘤。核酶特异性抑制VEGF 189可降低肺癌细胞系的血管分布和异种移植性。VEGF的细胞相关亚型（VEGF 189）可能在体内非小细胞肺癌异种移植物的间质血管化和生长中起关键作用。

项目成果

Okamoto K, Oshika Y, Fukushima Y, Ohnishi Y, Tokunaga T, Tomii Y, Kijima H, Yamazaki H, Ueyama Y, Nakamura M.: "Xenografts of human solid tumors frequently express cellular-associated isoform of vascular endothelial growth factor (VEGF) 189."Oncology Repo

Okamoto K、Oshika Y、Fukushima Y、Ohnishi Y、Tokunaga T、Tomii Y、Kijima H、Yamazaki H、Ueyama Y、Nakamura M.：“人类实体瘤的异种移植物经常表达细胞相关的血管内皮生长因子亚型（VEGF）

Tsuchida T, Kijima H, Oshika Y, Tokunaga T, Abe Y, Yamazaki H, Tamaoki N, Ueyama Y, Scanlon K J, Nakamura M: "Hammerhead ribozyme specifically inhibits mutant K-ras mRNA of human pancreatic cancer cells."Biochemical and Biophysical Research Communications

Tsuchida T、Kijima H、Oshika Y、Tokunaga T、Abe Y、Yamazaki H、Tamaoki N、Ueyama Y、Scanlon K J、Nakamura M：“锤头核酶特异性抑制人胰腺癌细胞的突变 K-ras mRNA。”生物化学和生物物理

Hatanaka H., Abe Y., Kamiya T., Morino F., Nagata J., Tokunaga T., Oshika Y., Suemizu H., Kijima H., Tsuchida T., Yamazaki H., Inoue H., Nakamura M.and Ueyama Y.: "Clinical implications of interleukin (IL)-10 induced by non-small-cell lung cancer."Annals

Hatanaka H.、Abe Y.、Kamiya T.、Morino F.、Nagata J.、Tokunaga T.、Oshika Y.、Suemizu H.、Kijima H.、Tsuchida T.、Yamazaki H.、Inoue H.、Nakamura M

Okamoto K, Oshika Y, Fukushima Y, Yoshimura M, Ohnishi Y, Tokunaga T, Hashimoto T, Hatanaka H, Tomii, Y, Yoshida Y, Miura S, Tsuchida T, Kijima H, Yamazaki H, Nakamura M, Ueyama Y.: "Inhibition of liver metastasis of colon cancer by in vivo administration

冈本 K、Oshika Y、福岛 Y、吉村 M、Ohnishi Y、德永 T、桥本 T、Hatanaka H、Tomii、Y、吉田 Y、三浦 S、土田 T、木岛 H、山崎 H、中村 M、上山 Y：

Tokunaga T, Nakamura M, Oshika Y, Abe Y, Ozeki Y, Fukushima Y, Hatanaka H, Sadahiro S, Kijima H, Tsuchida T, Yamazaki H, Tamaoki N, Ueyama Y.: "Thrombospondin 2 expression is correlated with inhibition of angiogenesis and metastasis of colon cancer."Briti

Tokunaga T, Nakamura M, Oshika Y, Abe Y, Ozeki Y, Fukushima Y, Hatanaka H, Sadahiro S, Kijima H, Tsuchida T, Yamazaki H, Tamaoki N, Ueyama Y.：“血小板反应蛋白 2 表达与血管生成抑制相关