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的细胞行为中的重要性的第一个线索来自对癌症的研究，癌症的特征是无序。纤维连接蛋白是正常细胞的主要细胞表面蛋白，而转化细胞表面不存在纤维连接蛋白。在一个对组织和细胞行为如此关键的网络中，非常不同的点上的缺陷都可能导致肿瘤细胞特有的紊乱，即细胞黏附丧失和细胞骨架紊乱。纤维连接蛋白是正常对照的癌细胞中的这些糖蛋白。我们注意到肿瘤与纤维连接蛋白的联系，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。我们建立了用兔抗…包被的胶乳测定尿中纤维连接蛋白浓度的新方法更多的是人类纤维连接蛋白。正常成人的尿纤维连接蛋白几乎为负值，但进展性胃肠癌患者的尿纤维连接蛋白水平显著高于非进展性胃肠癌患者。特别是，许多有肝转移和复发的病例会产生明确的、高水平的尿纤维连接蛋白，远高于无肝转移病例的水平。因此，由于胃肠癌和肝转移患者尿纤维连接蛋白浓度升高，尿纤维连接蛋白似乎是一种合适的肿瘤标记物。从胃肠道肿瘤患者尿液中提纯纤维连接蛋白，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。纯化胃癌和结肠癌患者尿液识别的约10万道尔顿的纤维连接蛋白片段。在肝细胞癌患者中，纯化其尿液识别的约100,000道尔顿和68,000道尔顿纤维连接蛋白片段。良性疾病患者血浆纤维连接蛋白和尿液纤维连接蛋白分别出现240000和200,000条道尔顿条带。然而，胃肠道肿瘤患者尿中的纤维连接蛋白是公认的降解片段，其相对分子质量为100,000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)，蛋白质的结合功能与其结构明显相关。为了确定胃肠道肿瘤患者尿纤维连接蛋白的结构域结构，我们用4株抗纤维连接蛋白结构域III、IV、VI的单抗对纯化的尿纤维连接蛋白进行了免疫印迹分析。较少

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