ADHESION PROTEIN AND OVARIAN CANCER IMPLANTATION

粘附蛋白和卵巢癌植入

• 批准号: 2101421
• 负责人: STEPHEN A CANNISTRA
• 金额: $ 20.03万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-08-01 至 1997-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2101421
• 关键词: CD antigens; athymic mouse; cell adhesion; cell adhesion molecules; clone cells; disease /disorder model; epithelium; gene expression; human subject; hyaluronate; laboratory mouse; ligands; mesothelioma; neoplasm /cancer invasiveness; neoplastic growth; neutralizing antibody; northern blottings; ovary neoplasms; peritoneum neoplasm; protein purification; proteins; southern blotting; transfection

Ovarian cancer is a highly lethal neoplasm due to its ability to form tumor implants on the peritoneal mesothelial surface of the abdominal cavity and bowel serosa. The mechanism by which ovarian cancer cells bind to peritoneal mesothelium most likely involves cell-cell recognition through specific adhesion molecules. In order to study this phenomenon, we have developed methods to isolate normal and malignant ovarian epithelial cells and have used an in vitro adhesion assay to quantitate the binding of these cells to peritoneal mesothelium. We have determined that the CD44 adhesion molecule is partly responsible for ovarian cancer cell binding through recognition of mesothelial-associated hyaluronate, a known ligand for CD44. In addition, we have found that ovarian cancer cells also bind through a CD44-independent mechanism which is not related to known adhesion proteins such as integrins or selectins. The first specific aim of this proposal is to determine the in vivo relevance of CD44 in the process of ovarian cancer cell implantation. By using a well-established nude mouse model, we will determine the effects of interfering with CD44 function (with neutralizing antibody or with soluble CD44 protein) on the implantation of CD44-positive ovarian cancer cells in vivo. We will also use a CD44-negative, implantation- incompetent ovarian cancer cell line to determine the effects of CD44 expression (by transfection) on in vivo implantation. In the second specific aim, the regulation of CD44 expression and function will be investigated in normal and malignant ovarian epithelial cells at the gene level using Southern and Northern analyses, and at the protein level using pulse-chase techniques. The third specific aim is to identify additional adhesion molecules which mediate the CD44-independent binding of ovarian cancer cells to peritoneal mesothelium. By using either ovarian cancer cells or mesothelial cells as immunogens, monoclonal antibodies will be generated and screened for their ability to neutralize binding. These neutralizing antibodies will provide useful tools for identifying potentially novel adhesion molecules expressed by either malignant ovarian epithelium or by peritoneal mesothelial cells. These studies should lead to a better understanding of the role of adhesion molecules in the process of ovarian cancer cell implantation. A complete characterization of these structures may eventually permit the design of adhesion antagonists capable of interfering with or reversing the intraabdominal spread of ovarian cancer.

卵巢癌是一种高度致命的肿瘤，因为它能够形成 肿瘤植入腹部腹膜间皮表面 腔和肠浆膜。 卵巢癌细胞的发生机制 与腹膜间皮的结合最有可能涉及细胞间识别 通过特定的粘附分子。 为了研究这一现象， 我们已经开发出分离正常和恶性卵巢的方法 上皮细胞并使用体外粘附测定来定量 这些细胞与腹膜间皮的结合。 我们已经确定 CD44粘附分子是卵巢癌的部分原因 通过识别间皮相关透明质酸盐进行细胞结合， CD44 的已知配体。 此外，我们还发现卵巢癌 细胞还通过不相关的 CD44 独立机制结合 已知的粘附蛋白，例如整合素或选择素。 该提案的第一个具体目标是确定体内 CD44在卵巢癌细胞植入过程中的相关性。 通过使用完善的裸鼠模型，我们将确定 干扰 CD44 功能的影响（使用中和抗体或 含可溶性 CD44 蛋白）对 CD44 阳性卵巢着床的影响 体内的癌细胞。 我们还将使用 CD44 阴性、植入- 无能卵巢癌细胞系以确定 CD44 的影响 体内植入时的表达（通过转染）。 在第二个 特定目标下，CD44表达和功能的调节将是 在正常和恶性卵巢上皮细胞中研究该基因 使用 Southern 和 Northern 分析以及蛋白质水平 使用脉冲追踪技术。 第三个具体目标是确定 介导不依赖 CD44 的结合的额外粘附分子 卵巢癌细胞转移至腹膜间皮。 通过使用 卵巢癌细胞或间皮细胞作为免疫原，单克隆 将产生抗体并筛选其中和能力 绑定。 这些中和抗体将为 识别潜在的新型粘附分子 恶性是由卵巢上皮或腹膜间皮细胞引起。 这些研究应该有助于更好地理解 卵巢癌细胞植入过程中的粘附分子。 这些结构的完整表征最终可能允许 能够干扰或逆转粘连拮抗剂的设计 卵巢癌的腹腔内扩散。