INTEGRIN RECEPTOR FOR LAMININ IN MALIGNANT MELANOMA

恶性黑色素瘤中层粘连蛋白的整合素受体

• 批准号: 3196592
• 负责人: RANDALL H KRAMER
• 金额: $ 15.29万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-04-01 至 1995-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3196592
• 关键词: affinity chromatography; antibody formation; basement membrane; cell adhesion; cell migration; cellular oncology; chemical binding; flow cytometry; human subject; human tissue; integrins; laboratory mouse; laminin; metastasis; neoplastic cell; nevus; posttranslational modifications; protein purification; protein structure function; receptor; receptor binding; receptor expression

Malignant melanoma cells must penetrate the basement membrane barrier both when they escape from the primary tumor and when they arrest in the microvasculature at distant sites. As tumor cells invade basement membrane, they interact with laminin, a major basement-membrane-specific glycoprotein that promotes cell attachment and migration. This interaction is mediated by multiple types of adhesion receptors. A novel integrin receptor complex that specifically binds laminin has recently been identified on human and mouse melanoma cells. This heterodimer complex has been tentatively designated alpha7beta1. The overall objectives of this proposal are to isolate and further characterize this unique melanoma- associated integrin, and to determine how this receptor modulates cell behavior on laminin and basement membrane substrates. The alpha7beta1 receptor complex will be purified from human melanoma cells by preparative ligand affinity chromatography for detailed immunochemical and biochemical analysis. The expression and distribution of the receptor in various tissues will be determined. Levels of the alpha7 receptor in normal and dysplastic nevi, and in primary and metastatic melanoma will be compared in order to establish a possible link between receptor expression and tumor progression. The distinctive alpha subunit will be compared with the other known integrin beta1-associated alpha subunits by peptide mapping and N- terminal amino acid sequence analysis. How the alpha7 and beta1 subunits are processed by post-translational modifications and when ligand binding activity is acquired will be followed. Studies with isolated subdomains of laminin will identify the specific sites that bind the alpha7 receptor. Antibodies that block receptor function will be developed and tested for their effects on adhesion, migration, and invasion. Integrin receptor profiles will be compared in sets of established human and mouse melanoma cell lines with high and low metastatic potential. Panels of variant tumor cell lines with altered receptor profiles will be selected from parental cell populations by flow cytometry and tested for their invasive behavior. These studies should increase our understanding of how laminin-binding receptors, such as the novel alpha7 complex, influence the metastatic behavior of malignant melanoma cells.

恶性黑色素瘤细胞必须穿透基底膜屏障 当他们从原发性肿瘤中逃脱时 远处的微脉管系统。 随着肿瘤细胞入侵地下室 膜，它们与层粘连蛋白相互作用，层粘连蛋白是一个主要的地下室膜特异性 促进细胞附着和迁移的糖蛋白。 这种相互作用 由多种类型的粘附受体介导。 一种新颖的整合素 特异性结合层粘连蛋白的受体复合物最近已经 在人类和小鼠黑色素瘤细胞上鉴定。 这个异二聚体复合物具有 被指定为alpha7beta1。 总体目标 建议是分离并进一步表征这种独特的黑色素瘤 相关的整合素，并确定该受体如何调节细胞 层粘连蛋白和基底膜底物的行为。 alpha7beta1 受体复合物将通过制备者从人黑色素瘤细胞中纯化 配体亲和色谱法，用于详细的免疫化学和生化 分析。 受体在各种中的表达和分布 将确定组织。 正常和 将比较发育不良的NEVI，以及在原发性和转移性黑色素瘤中 为了在受体表达与肿瘤之间建立可能的联系 进展。 将独特的α亚基与其他亚基进行比较 已知的整合素β1相关α亚基通过肽映射和N- 末端氨基酸序列分析。 alpha7和beta1亚基如何 通过翻译后修饰和配体结合时处理 将遵循活动。 研究的研究 层粘连蛋白将识别结合α7受体的特定位点。 将开发并测试阻断受体功能的抗体 它们对粘附，迁移和入侵的影响。 整联蛋白受体 将在成熟的人类和小鼠黑色素瘤组中比较曲线 具有高转移势和低转移性的细胞系。 var 将从父母中选择具有改变受体轮廓的细胞系 通过流式细胞仪进行细胞群体，并测试了其侵入性行为。 这些研究应该增加我们对层粘连蛋白结合的理解 受体，例如新型α7复合物，会影响转移性 恶性黑色素瘤细胞的行为。