Development of anti-cancer reagent utilizing cytotoxic potential of human ribonucleases

利用人类核糖核酸酶的细胞毒性潜力开发抗癌试剂

• 批准号: 09555255
• 负责人: YAMADA Hidenori
• 金额: $ 6.78万
• 依托单位: Okayama University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09555255/
• 关键词: RNase; Anti-cancer agent; Growth factor; Growth inhibition; 細胞成長因子; 細胞増殖阻害

Pancreatic-type RNases are considered to have cytotoxic potential, because they can degrade RNA molecules when enter the cytosol. However, most of these RNases are virtually little toxic because cells have no active uptake mechanism for these RNases and because ubiquitous cytoplasmic RNase inhibitor is considered to play protective role against endocytotic leak of RNases from outside of cells. To study the cytotoxic potential of RNase toward malignant cells targeting growth factor receptors, the carboxyl terminus of human RNase 1 was fused to the amino terminus of human basic fibroblast growth factor (bFGF). This RNase-FGF fused protein was found to effectively inhibited the growth of mouse melanoma cell line B 16/BL6 with the high level of cell surface FGF receptor.This effect appeared to result from prolongation of overall cell cycle rather than killing cells or specific arrest in a particular phase of cell cycle. Thus human RNase 1 fused to a ligand of cell surface molecules such as FGF receptor is shown to be an effective candidate for selective cell targeting agent toward malignant cells with low toxic effects on normal cell types.

胰腺型 RNase 被认为具有细胞毒性潜力，因为它们在进入细胞质时可以降解 RNA 分子。然而，大多数这些 RNase 实际上毒性很小，因为细胞对这些 RNase 没有主动摄取机制，并且普遍存在的细胞质 RNase 抑制剂被认为对防止 RNase 从细胞外的内吞渗漏发挥保护作用。为了研究 RNase 对靶向生长因子受体的恶性细胞的细胞毒性潜力，将人 RNase 1 的羧基末端与人碱性成纤维细胞生长因子 (bFGF) 的氨基末端融合。这种 RNase-FGF 融合蛋白被发现能有效抑制具有高水平细胞表面 FGF 受体的小鼠黑色素瘤细胞系 B 16/BL6 的生长。这种效应似乎是由于整个细胞周期的延长而不是杀死细胞或在细胞周期的特定阶段进行特异性停滞。因此，与细胞表面分子（例如 FGF 受体）的配体融合的人 RNase 1 被证明是针对恶性细胞的选择性细胞靶向剂的有效候选者，并且对正常细胞类型具有低毒性作用。

项目成果

J.Futami: "Tissue specific expression of pancreatic-type RNases and RNase inhibitor in humans." DNA and Cell Biology. 16・4. 413-419 (1997)

J.Futami：“人类胰腺型 RNase 和 RNase 抑制剂的组织特异性表达。”16·4（1997）。

M.Seno: "BALB/C 3TC Cells Co-Expressing FGF-2 and soluble FGF Receptor Acquire Tumorigenecity." Cytokine. (発表予定).

M.Seno：“BALB/C 3TC 细胞共表达 FGF-2 和可溶性 FGF 受体获得细胞因子。”

T.E.Creighton(Editor): "Protein Function;A Practical Approach" IRL Press (at Oxford University Press,Oxford,New York,Tokyo), 334 (1997)

T.E.Creighton（编辑）：“蛋白质功能；实用方法”IRL Press（牛津大学出版社，牛津，纽约，东京），334（1997）

M.Ueda: "Molecular Targeting for Epidermal Growth Factor Receptor Expressed on Breast Cancer Cells." Breast Cancer. 4・4. 67-69 (1997)

M.Ueda：“乳腺癌细胞上表达的表皮生长因子受体的分子靶向”，4·4（1997）。

S.S.Terzyn: "The three dimensional structure of human RNase4, unliganded and complexed with d(Up), reveals the basis for its uridine selectivity." Journal of Molecular Biology. 285.1. 205-214 (1999)

S.S.Terzyn：“人类 RNase4 的三维结构（未配体并与 d(Up) 复合）揭示了其尿苷选择性的基础。”