TISSUE FACTOR MESSENGER RNA, ANTIGEN, AND ACTIVITY

本申请的主要目的是评估 组织因子（TF）的合成及表达 癌细胞中的促凝血活性，以确定 TF在恶性肿瘤相关凝血病中的作用以及 TF在肿瘤细胞中的增殖能力。 的潜在 抑制TF或TF依赖性凝血系统的益处 对于治疗益处和早期转移灶的诊断， 被评价。 我们将使用的具体目标和方法来实现 目的：1）TF信使RNA（mRNA），TF 抗原和TF活性将在各种人类肿瘤中测定。 体外细胞系。 这将提供基线信息 关于TF基因表达和生物活性。 这一目标 将通过RNA分离和北方印迹分析完成 使用先前已开发的cDNA探针;单克隆 将在ELISA试验中使用抗体检测TF抗原; TF 促凝血活性将通过使用两阶段 凝血试验，将用纯的 人脑TF 2)各种试剂，如生长因子， 化学治疗剂或炎症介质，其 已知在正常细胞中刺激或抑制TF活性 （成纤维细胞、内皮细胞、单核细胞等），将 评估了对TF mRNA、TF抗原和TF活性的影响， 肿瘤细胞系 这些信息将作为基线， 癌细胞对各种药物的TF反应。 3)TF mRNA，TF 抗原和TF活性以及这些变量在 将在恶性细胞中评价对各种试剂的反应 来源于与细胞系相同的细胞类型，但获得 从病人身上。 4)肿瘤组织TF mRNA、TF抗原和TF活性 细胞将在注射后进行体内评价， 在裸鼠中形成。 5)对TF mRNA、TF抗原和 抑制或刺激TF的各种试剂的TF活性将被测定。 在裸鼠体内评价以及各种试剂的作用 对肿瘤发生和存活率的影响。 对致瘤性的影响将 通过肿瘤大小和组织病理学分析进行评价， 与标记TF单克隆抗体的诊断成像相关 抗体的 本项目将提供TF基因的相关性 表达，无论是转录和翻译与外观 在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， 恶性肿瘤中TF与肿瘤发生率增加的相关性 与那些具有较低致瘤潜力的人相比。

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.