HUMAN BASEMENT MEMBRANE TUMORS

基底膜是重要的细胞外结构，与 细胞黏附、生长、分化和癌症转移。这个 基膜的主要结构成分，包括IV型 胶原蛋白、层粘连蛋白、纤维连接蛋白、蛋白多糖和牙本质蛋白 目前正在对它们的具体功能进行调查。 这些研究的基膜组分主要是衍生出来的 来自小鼠(EHS肉瘤)或大鼠(卵黄囊肿瘤)，因为有 一直不是人类基底膜的良好来源。主要目标 该项目的目的是建立人的基底膜，使其在 细胞培养和裸鼠作为人类的永久来源 基底膜成分。该项目的第二个目标是 人源性基底膜的结构和功能比较 与它们的小鼠衍生的对应物。人类肿瘤与 最高基底膜生产能力将通过以下方式确定 使用IV型抗体的组织学和免疫细胞化学方法 胶原蛋白和层粘蛋白。初步筛查表明，人类 基底膜合成量最高的肿瘤包括 涎腺腺样囊性癌，卵黄囊(内胚层) 睾丸肿瘤和肾腺癌。脑部肿瘤 上述类别具有最高的免疫细胞化学演示 在细胞培养和AS中都将建立基底膜 裸鼠的异种移植。层粘连蛋白和IV型胶原将是 从人类肿瘤中提取的方法与提取时使用的方法相同 死于EHS肉瘤。这些成分的合成速度将是 用C14脯氨酸和S35蛋氨酸放射性标记测定 这将与EHS肉瘤的合成率进行比较。这个 人体肿瘤基底膜的产生能力将 随着时间的推移通过细胞传代和异种移植进行监测。这个 人层粘连蛋白和IV型胶原的结构和功能 与小鼠的SDS-PAGE、肽图谱比较， 旋转阴影、附着试验和抗体交叉激活。

Basement membranes are important extracellular structures implicated in cellular adhesion, growth, differentiation, and cancer metastasis. The main structural components of basement membranes including Type IV collagen, laminin, fibronectin, proteoglycan, and entactin have been identified and their specific functions are now being investigated. Basement membrane components for these studies have been derived mainly from murine (EHS sarcoma) or rat (yolk sac tumor) sources, because there has not been a good source of human basement membranes. The main objective of this project is to establish human basement membrane producing tumors in both cell culture and in athymic (nude) mice as a perpetual source of human basement membrane components. A second objective of this project is to compare the structure and function of the human-derived basement membrane components with their murine-derived counterparts. Human tumors with the highest basement membrane producing capability will be identified by histologic and immunocytochemical methods using antibodies to Type IV collagen and laminin. Preliminary screening has indicated that the human tumors with the highest amounts of basement membrane synthesis include the adenoid cystic carcinoma of the salivary gland, the yolk sac (endodermal sinus) tumor of the testis, and adenocarcinoma of the kidney. Tumors of the above categories with the highest immunocytochemical demonstration of basement membrane will be established in both cell culture and as xenografts in athymic "nude" mice. Laminin and Type IV collagen will be extracted from the human tumors by identical methods used in the extraction from the EHS sarcoma. The rate of synthesis of these components will be determined by using radiolabeling with C14 proline and S35 methionine and this will be compared to the synthesis rate of the EHS sarcoma. The basement membrane producing capability of the human tumors will be monitored over time with cell passage and xenograft transplantation. The structure and function of human laminin and Type IV collagen will be compared to their murine counterparts with SDS-PAGE, peptide mapping, rotary shadowing, attachment assays, and antibody cross-reactivation.