ACBT GLIOMA SPHEROIDS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 急性、慢性和重复PDT后的ACBT球状体将成像用于活-死测定和细胞凋亡。 基因治疗的治疗有效性通常受到大分子向胞质溶胶中的低效转移以及缺乏位点特异性靶向的限制。光化学内化（PCI）是基于光化学诱导的内吞大分子从内体和溶酶体释放到胞质溶胶中。PCI已与基因治疗联合使用，并已显示增加基因表达。将在体内和体外研究PCI增强肿瘤抑制基因PAX 6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RG 2的单层和多细胞球体培养物将用于体外研究。相同的细胞系将用于诱导大鼠颅内肿瘤。共聚焦和多光子荧光显微镜将用于研究光敏剂分布，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. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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.