ACBT GLIOMA SPHEROIDS

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 急性、慢性和重复PDT后的ACBT球体将被成像用于活体死亡分析和细胞凋亡。 基因治疗的有效性往往受到大分子向胞浆的低效转移以及缺乏定点靶向的限制。光化学内化是基于光化学诱导内吞大分子从胞内体和溶酶体释放到胞浆中的过程。经皮冠状动脉介入治疗已与基因治疗结合使用，并已被证明可提高基因表达。我们将在体内和体外研究经皮冠状动脉介入术促进肿瘤抑制基因PAX6或报告基因插入胶质瘤细胞的能力。大鼠胶质瘤细胞系F98和RG2的单层和多细胞球体培养将用于体外研究。同样的细胞系将被用来诱导大鼠的颅内肿瘤。共聚焦和多光子荧光显微镜将用于研究光敏剂的分布、报告基因(GFP)的表达和细胞存活。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. ACBT spheroids following acute, chronic, and repetitive PDT will be imaged for live-dead assays and apoptosis. The therapeutic usefulness of gene therapy is often limited by an inefficient transfer of the macromolecule into the cytosol, and a lack of site-specific targeting. Photochemical internalization (PCI) is based on photochemically induced release of endocytosed macromolecules from endosomes and lysosomes into the cytosol. PCI has been used in combination with gene therapy and has been shown to increase gene gene expression. The ability of PCI to enhance the gene insertion of the tumor suppressor gene PAX6 or reporter genes into glioma cells will be studied in vivo and in vitro. Monolayer and multicellular spheroid cultures of the rat glioma cell lines F98 and RG2 will be used in the in vitro studies. The same cell lines will be used to induce intracranial tumors in rats. Confocal and multi photon fluorescence microscopy will be used to study photosensitizer distribution, expression of reporter gene (GFP), and cell survival.