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（VEGF189）的细胞相关同工型的具体作用尚未澄清，而五种具有不同生物学活性的VEGF的同工型（VEGF121，VEGF145，VEGF145，VEGF165，VEGF165，VEGGF165，VEGF189，VEGGF206）是通过替代性生成的。我们使用了锤头型核酶（V189RZ）抑制VEGF189 mRNA。v189rz特异性裂解的VEGF189 mRNA的外显子6，但没有针对VEGF121或VEGF165同工型的活性。 V189RZ被引入到人类非小细胞肺癌细胞系（OZ-6/VR）中。在OZ-6/VR中，VEGF189 mRNA的表达水平降低，而VEGF121和165表达则保留了。 OZ-6/VR异种移植被认为显着降低了血管化和生长，而细胞系在组织培养条件下没有显示出任何改善的生长。 OZ-6/VR细胞（1×10^5/小鼠）没有肿瘤，而亲本的OZ-6细胞在8周内形成了巨大的肿瘤。核酶对VEGF189的特异性抑制降低了肺癌细胞系的血管性和异种植物性。 VEGF（VEGF189）与细胞相关的同工型可能在体内非小细胞肺癌Xenographictics的基质血管化和生长中具有关键作用。

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

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

Tomisawa M, Tokunaga T, Oshika Y, Tsuchida T, Fukushima H, Sato H,Kijima H, Yamazaki H, Ueyama Y, Tamaoki N, Nakamura M.: "Expression pattern of vascular endothelial growth factor isoform is closely correlated with tumour stage and vascularization in rena

Tomisawa M、Tokunaga T、Oshika Y、Tsuchida T、Fukushima H、Sato H、Kijima H、Yamazaki H、Ueyama Y、Tamaoki N、Nakamura M.：“血管内皮生长因子亚型的表达模式与肿瘤分期和肿瘤分期密切相关。