Purification of abnormal fibronectin fragment in urine of cancer patient and development of gastrointestinal cancer diagnosis with it's monoclonal antibody.

纤维连接蛋白存在于正常人体组织和基底膜中。纤维连接蛋白在有组织细胞行为中的重要性的第一个线索来自癌症的研究，癌症的特征是紊乱。纤维连接蛋白是正常细胞的主要细胞表面蛋白，但在转化细胞表面不存在。在一个对组织和细胞行为至关重要的联系中，在不同的点上的缺陷都可能导致肿瘤细胞的特征性紊乱，即细胞粘附的丧失和细胞骨架的紊乱。纤维连接蛋白被认为是癌细胞中正常对照的这些糖蛋白。我们注意到癌症与纤维连接蛋白的联系，并且似乎与细胞外基质纤维连接蛋白的降解有关，当癌细胞继续增殖和转移时，其降解产物出现在尿液中。我们建立了一种新的测定尿中纤维连接蛋白浓度的方法，用乳胶涂覆兔抗…正常成人的尿纤维连接蛋白值几乎为阴性，但进展性胃肠道癌患者的尿纤维连接蛋白值明显高于未患这种疾病的患者。特别是，许多肝转移和复发的病例产生明确的、高水平的尿纤维连接蛋白，远高于没有肝转移的病例。基于这些原因，并且由于尿纤维连接蛋白浓度升高在胃肠道癌和肝转移中发现，尿纤维连接蛋白似乎是一个合适的肿瘤标志物。从胃肠道肿瘤患者尿液中纯化纤维连接蛋白，并与血浆纤维连接蛋白和良性疾病患者尿液中的纤维连接蛋白进行比较。纯化胃癌、结肠癌患者尿液中识别的约10万道尔顿纤维连接蛋白片段。在肝细胞癌患者中，净化尿液中识别的约100,000道尔顿和68,000道尔顿的纤维连接蛋白片段。SDS-PAGE结果显示，高血压患者血浆纤维连接蛋白存在24万道尔顿带，尿纤维连接蛋白存在20万道尔顿带。然而，在胃肠道肿瘤患者的尿中，纤维连接蛋白是公认的降解片段，其分子量为10万道尔顿。纤维连接蛋白分子首先被分解成离散的蛋白质结构域（结构域I-VII），蛋白质的结合功能及其结构明显相关。为了确定胃肠道肿瘤患者尿纤维连接蛋白结构域的结构，我们使用4种针对纤维连接蛋白结构域III、IV和VI的单克隆抗体对纯化的尿纤维连接蛋白进行了westwen blot分析。少

The distribution of fibronectin exists in normal human tissue and basement membrane.The first clues to the importance of fibronectin in organized cell behavior came from studies of cancer, which is characterized by disorder. Fibronectin is a major cell surface protein of normal cells but is absent from the surface of transformed cells. Within a nexus so crucial to organization and cell behavior, defects at very different points could all lead to the characteristic disarray of tumor cells, that is the loss of cell adhesion and the disorderd cytoskeleton. The fibronectins are the understood of these glvcoproteins of normal controls in cancer cells. We attending to connect of cancers and fibronectin, and would seem to bear a relation to a degrade extracellular matrix fibronectins, when the cancer cells went on multiplying and metastasis, and its the degrade products appeared in urine. We were developed new method from fibronectin concentration in urine, using latex coated with rabbit anti … More human fibronectin. Urine fibronectin values in normal adults were almost negative, but were significantly higher in patients with progressive gastrointestinal cancer than those not so afflicted. Especially, many cases of liver metastasis and recurrence produced well-defined, high levels of urine fibronectin, well above levels in cases whithout liver metastasis. From these reasons, and because elevated urine fibronectin concentration is found in gastrointestinal cancer and liver metastasis, urine fibronectin would seem to be a suitable tumor marker. Fibronectin was purified from urine of patient with gastrointestinal cancers were compared with that of plasma fibronectin and that of patients urine with benign diseases. Purifying the about 100,000-dalton fibronectin fragment recognized by gastric cancer and colon cancer patients urine. In hepatocellular carcinoma patients, purifying the about 100,000-dalton and 68,000-dalton fibronectin fragments recognized by their urine. According to SDS-PAGE, 240000-dalton band was present in plasma fibronectin and 200,000 dalton band was present in urine fibronectin of benigh disease. However, fibronectin in gastrointestinal cancer patients urin, which MW is 100,000-dalton, was recognized fragment of degradation. The fibronectin molecule was dissected first into discrete protein domains(domain I-VII) and theprotein's binding functions and its structure are clearly correlated. To determine the domain structure of urinary fibronectin in gastrointestinal cancer patients, we carried out the westwen blot analysis of the purified urinary fibronectins by used with four monoclonal antibodies to domain III,IV, and VI of fibronectin. Less