Molekulare Charakterisierung von Glutamat-Rezeptoren in humanen Tumoren

谷氨酸n -甲基- d -天冬氨酸（NMDA）和¿-氨基-3-羟基-5-甲基-4-异恶唑丙酸（AMPA）受体拮抗剂主要用于神经系统适应症，在体外和体内对多种人类癌症具有有效的抗癌作用。在本项目的背景下，我们计划通过RT-PCR分析，免疫细胞化学和共聚焦显微镜研究NMDA和AMPA受体亚基在神经组织中广泛表征的各种人类癌症中的表达和细胞内定位。我们将通过慢病毒载体的RNA干扰来阻断不同亚基在癌细胞增殖、迁移和存活中的表达，从而研究不同亚基在癌细胞增殖、迁移和存活中的功能意义。此外，我们希望利用免疫沉淀结合SDS-PAGE鉴定与癌细胞中谷氨酸受体亚基相互作用的蛋白伴侣。分离的相互作用蛋白候选伙伴随后将使用基质辅助激光解吸/电离质谱（MALDI-MS）进行序列分析。

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).