Identification of porin specifically expressed in tumor cells and its application for development of anti-tumor drugs

肿瘤细胞特异性表达孔蛋白的鉴定及其在抗肿瘤药物开发中的应用

• 批准号: 08672560
• 负责人: SHINOHARA Yasuo
• 金额: $ 1.54万
• 依托单位: The University of Tokushima
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08672560/
• 关键词: Hexokinase; porin; cDNA cloning; tumor cells; energy metabolism; anti tumor drugs; VDAC; ミトコンドリア

First of all, to understand the physiological meanings of hexokinase binding to mitochondria observed in tumor cells, we isolated and characterized tumor mitochondria. As a result, mitochondria-bound hexokinase was found to use mitochondrial ATP preferentially than thatsynthesized in cytosol. Furthermore, this binding was found to enable the efficient cross talk between glycolysis and oxydative phosphorylation.Next, to understand the molecular mechanisms of hexokinase binding to mitocondrial membrane, we tried to identify the porin isoform (s) expressed as binding site (s) of hexokinase in tumor mitochondria. As a result,1) we succeeded to isolate two kinds of cDNA clones expressed in AH130 cells2) these cDNA clones were found to encode two kinds of porin isoform called as VDAC1 and VDAC23) furthermore, both of VDAC1 and VDAC2 were found to be expressed in mitochondrial membrane.

首先，为了理解在肿瘤细胞中观察到的己糖激酶与线粒体结合的生理意义，我们分离并表征了肿瘤线粒体。结果发现，与胞浆中合成的ATP相比，线粒体结合的己糖激酶优先使用线粒体ATP。此外，这种结合被发现能够使糖酵解和氧化磷酸化之间的有效串扰。接下来，为了理解己糖激酶与线粒体膜结合的分子机制，我们试图鉴定肿瘤线粒体中作为己糖激酶结合位点表达的孔蛋白同种型。结果表明：1）成功分离到两种在AH 130细胞中表达的cDNA克隆; 2）这些cDNA克隆编码两种孔蛋白亚型VDAC 1和VDAC 23;

项目成果

Y.Shinohara, et al.: "Source of ATP for hexokinase-catalyzed glucose phosphorylation in tumor cells : Dependence on the rate of oxidative phosphorylation to that of extramitochondrial ATP generation."Biochim.Biophys.Acta. 1319. 319-330 (1997)

Y.Shinohara 等人：“肿瘤细胞中己糖激酶催化葡萄糖磷酸化的 ATP 来源：氧化磷酸化速率与线粒体外 ATP 生成速率的依赖性。”Biochim.Biophys.Acta。

篠原 康雄、寺田 弘: "がん細胞のエネルギー代謝系のキャラクタリゼーション" サイエンスリポート 印刷中 (総説).

Yasuo Shinohara、Hiroshi Terada：“癌细胞能量代谢系统的特征”科学报告已出版（评论）。

Y.Shinohara et al.: "Quantitative determinations of the steady state transcript levels of hexokinase isozymes and glucose transporter isoforms in normal rat tissues and the malignant tumor cell line AH130"Biochim.Biophys.Acta. 1368. 129-136 (1998)

Y.Shinohara 等人：“正常大鼠组织和恶性肿瘤细胞系 AH130 中己糖激酶同工酶和葡萄糖转运蛋白同工型的稳态转录水平的定量测定”Biochim.Biophys.Acta。

篠原康雄、寺田 弘: "がん細胞のエネルギー代謝系のキャラクタリゼーション" サイエンスリポート 印刷中 (総説).

Yasuo Shinohara、Hiroshi Terada：“癌细胞能量代谢系统的特征”科学报告已出版（评论）。

Y.Shinohara et al.: "Quantitative determinations of the steady state transcript levels of hexockinase isozymes and glucose transporter isoforms in normal rat tissues and the malignant tumor cell line AH130" Biochim.Biophys.Acta. 1368. 129-136 (1998)

Y.Shinohara 等人：“正常大鼠组织和恶性肿瘤细胞系 AH130 中己糖激酶同工酶和葡萄糖转运蛋白同工型的稳态转录水平的定量测定”Biochim.Biophys.Acta。