TISSUE FACTOR MESSENGER RNA, ANTIGEN, AND ACTIVITY

此应用程序的主要目标是评估 组织因子(TF)的合成及其表达 癌细胞中促凝血剂的活性以确定 转铁蛋白在恶性肿瘤相关凝血疾病中的作用及其机制 Tf对肿瘤细胞增殖能力的影响。潜力 抑制组织因子或依赖于组织因子的凝血系统的益处 对于早期转移灶的治疗效益和诊断将 被评估。我们将用来实现的具体目标和方法 这些目标如下：1)转铁蛋白信使核糖核酸(MRNA)，转铁蛋白 抗原和转铁蛋白活性将在各种人类肿瘤中进行测定 体外培养的细胞系。这将提供基线信息 关于转铁蛋白基因的表达和生物活性。这个目标 将通过RNA分离和Northern印迹分析来完成 使用先前开发的cDNA探针的；单克隆性的 抗体将用于酶联免疫吸附试验以检测转铁蛋白抗原 促凝血剂的活性将通过两个阶段进行评估 凝血试验将用纯净的样品校准 人脑转运蛋白。2)各种制剂，如生长因子， 化疗药物，或炎症介质，它们是 已知可刺激或抑制正常细胞中的转铁蛋白活性 (成纤维细胞、内皮细胞、单核细胞等) 评估对Tf mRNA、Tf抗原和Tf活性的影响 肿瘤细胞系。此信息将作为基准 癌细胞中的转铁蛋白对各种药物的反应。3)转铁蛋白mRNA、转铁蛋白 抗原、转铁蛋白活性及这些变量的变化 恶性细胞对不同药物的反应将被评估 起源于与细胞系相同的细胞类型，但获得 从病人那里。4)肿瘤组织中的转铁蛋白mRNA、转铁蛋白抗原和转铁蛋白活性 注射和肿瘤后将在体内对细胞进行评估 在裸鼠体内形成。5)对转铁蛋白mRNA、转铁蛋白抗原及 抑制或刺激转铁蛋白的各种制剂的转铁蛋白活性 裸鼠体内评价及多种药剂的作用 关于致瘤性和生存性。对致瘤性的影响将 通过肿瘤大小和组织病理学分析和意志来评估 与标记转铁蛋白单抗的诊断成像相关 抗体。该项目将提供转铁蛋白基因的相关性 表达，既有转录，也有翻译，有外观 已知的恶性肿瘤中的生物活性 与血栓形成增加相关或不相关 转铁蛋白与恶性肿瘤致瘤性增高的相关性研究 潜在致癌潜力与致癌潜力较小的人。

The major objectives of this application are to evaluate the synthesis of tissue factor (TF) and the expression of TF procoagulant activity in cancer cells in order to determine the role of TF in malignancy-associated coagulapathies and the role of TF in the proliferative capacity of tumor cells. The potential benefit of inhibition of TF or the TF-dependent coagulation system for therapeutic benefit and diagnosis of early metastatic foci will be evaluated. The specific aims and methods we will use to achieve these objectives are as follows: 1) TF messenger RNA (mRNA), TF antigen, and TF activity will be determined in various human tumor cell lines in vitro. This will provide baseline information regarding TF gene expression and biological activity. This goal will be accomplished by RNA isolation and Northern blot analysis using cDNA probes which have previously been developed; monoclonal antibody will be used in an ELISA assay to detect TF antigen; TF procoagulant activity will be evaluated by use of a two-stage coagulation assay which will be calibrated with a sample of pure human brain TF. 2) Various agents, such as growth factors, chemotherapeutic agents, or mediators of inflammation, which are known to stimulate or inhibit TF activity in normal cells (fibroblasts, endothelial cells, monocytes, etc.), will be evaluated for effect on TF mRNA, TF antigen, and TF activity in tumor cell lines. This information will serve as a baseline for TF response in cancer cells to various agents. 3) TF mRNA, TF antigen, and TF activity and alteration of these variables in response to various agents will be evaluated in malignant cells originating from the same cell type as the cell lines, but obtained from patients. 4) TF mRNA, TF antigen, and TF activity of tumor cells will be evaluated in vivo following injection and tumor formation in nude mice. 5) The effect on TF mRNA, TF antigen, and TF activity of various agents that inhibit or stimulate TF will be evaluated in vivo in nude mice as well as effect of various agents on tumorigenicity and on survival. Effect on tumorigenicity will be evaluated by tumor size and by histopathologic analysis and will be correlated with diagnostic imaging of labelled TF monoclonal antibody. This project will provide correlation of TF gene expression, both transcription and translation with the appearance of biological activity in malignancies known to be either associated or not associated with increased thrombosis and correlation of TF in malignancies with increased tumorigenic potential versus those with less tumorigenic potential.