MAINTENANCE AND REPAIR OF VASCULAR ENDOTHELIUM

我们的具体目标是分析转染血管内皮细胞是否 bFGF cDNA表达载体转染内皮细胞后， 血管内皮细胞的自主增殖及其 转化成肿瘤细胞。 我们先来分析一下 bFGF是否会在细胞内起作用（如大多数癌基因 产品）或需要由细胞释放， 与暴露的细胞表面受体相互作用。 我们还将分析 它的细胞定位，以确定它是否可以 最终与细胞核、细胞质或细胞浆结合 膜分数 由于细胞转化可以是 癌基因表达的结果，我们将分析转染 血管内皮细胞是否组成性表达c 可见fos、c-myc和c-ras癌基因。 是否 bFGF基因产物可从联合细胞中释放出来 与ECM组分如硫酸乙酰肝素GAG接触， 有丝分裂原应具有高亲和力，将进行评估。 效果 TGFB和TNF对亲本细胞和转染细胞生长的影响 以及PA表达，将进行分析，以便 确定它们是否会产生类似于 当细胞在固体基质上生长时报告。 单克隆 将获得针对FGF受体的抗体并用于 开发FGF受体的灵敏RIA，以及 研究其表达方式。 它们也将用于分离FGF 通过免疫亲和色谱法测定受体。fgf受体 然后将在结构上进行表征， 获得的将用于开发特异性寡核苷酸探针， 其将用于2 FGF受体cDNA的克隆。

Our specific aims are to analyze whether transfection of vascular endothelial cells with bFGF cDNA expression vector could lead to autonomous proliferation of vascular endothelial cells and their transformation into neoplastic cells. We will first analyze whether bFGF would act intracellularly (as most oncogene products do) or need to be released by the cells in order to interact with exposed cell surface receptors. We will also analyze its cellular localization, in order to determine whether it could ultimately become associated with the nuclear, cytosol, or plasma membrane fraction. Since cell transformation can be the direct result of oncogene expression, we will analyze in transfected vascular endothelial cells whether constitutive expression of c fos, c myc and c ras oncogenes can be observed. Whether the bFGF gene product could be released from the cells in association with ECM components such as heparan sulfate GAG, for which the mitogen should have a high affinity, will be assessed. The effect of TGFB and TNF on the growth of parental and transfected cells in soft agar, as well as PA expression, will be analyzed in order to determine whether they could have effects similar to those reported when cells are grown on solid substrates. Monoclonal antibodies against the FGF receptor will be obtained and used for the development of sensitive RIA for the FGF receptor, and studies of its expression. They will also be used to isolate FGF receptors by immunoaffinity chromatography. The FGF receptor will then be characterized structurally, and the information gained will be used to develop sepecific oligonucleotide probes, which will be used for the cloning of 2 FGF receptor cDNA.