Molekulare Charakterisierung von Glutamat-Rezeptoren in humanen Tumoren

人类肿瘤中谷氨酸受体的分子特征

• 批准号: 25249853
• 负责人: Professorin Dr. Hrissanthi Ikonomidou
• 金额: --
• 依托单位: University of Wisconsin-Madison
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2006
• 资助国家: 德国
• 起止时间: 2005-12-31 至 2009-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/25249853?language=en
• 关键词: Molekulare; Charakterisierung; von; Glutamat; Rezeptoren

Glutamate N-methyl-D-aspartate (NMDA) and ¿-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptor antagonists, which have been developed primarily for neurological indications, elicit potent anticancer effects in a variety of human cancers in vitro and in vivo. In the context of this project we plan to investigate the expression and intracellular localization of NMDA and AMPA receptor subunits, which have been extensively characterized in neuronal tissue, in various human cancers by means of RT-PCR analysis, immunocytochemistry and confocal microscopy. We will examine functional significance of the different subunits for cancer cell proliferation, migration and survival by blocking their expression by means of RNA interference using lentiviral vectors. Furthermore, we want to identify protein partners which interact with glutamate receptor subunits in cancer cells using immunoprecipitation combined with SDS-PAGE. Isolated interaction protein partner candidates will be subsequently subjected to sequence analysis using matrix-assisted laser desorption/ionization mass spectroscopy (MALDI-MS).

谷氨酸N-甲基-D-天冬氨酸（NMDA）和â-氨基-3-羟基-5-甲基-4-异恶唑丙酸酯（AMPA）受体拮抗剂主要针对神经系统适应症开发，在多种人类癌症中产生强效抗癌作用体外和体内。在这个项目的背景下，我们计划调查的表达和细胞内定位的NMDA和AMPA受体亚基，这已被广泛的特点，在神经组织中，在各种人类癌症的RT-PCR分析，免疫细胞化学和共聚焦显微镜。我们将通过使用慢病毒载体通过RNA干扰阻断它们的表达来检查不同亚基对癌细胞增殖、迁移和存活的功能意义。此外，我们希望通过免疫沉淀结合SDS-PAGE鉴定与癌细胞中谷氨酸受体亚基相互作用的蛋白质伴侣。随后使用基质辅助激光解吸/电离质谱法（MALDI-MS）对分离的相互作用蛋白伴侣候选物进行序列分析。