DETECTION OF ALTERED APC PROTEINS IN COLON CANCER CELLS

结肠癌细胞中 APC 蛋白改变的检测

• 批准号: 3493423
• 负责人: David E. Hill
• 金额: $ 5万
• 依托单位: OMNIGENE DIAGNOSTICS, INC.
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-04-01 至 1993-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3493423
• 关键词: Chordata; colorectal neoplasms; diagnosis design /evaluation; gene mutation; human subject; immunofluorescence technique; immunologic assay /test; immunoprecipitation; neoplasm /cancer immunodiagnosis; neoplastic cell; prognosis; tissue /cell culture; tumor antigens; western blottings

Mutations in the APC gene appear to be crucial for adenoma formation in inherited Familial Adenomatous Polyposis (FAP) and in the early stages of colorectal cancer. Analysis of the APC gene has identified mutations in germ line DNA of FAP patients. Over 90% of the mutations interrupt the reading frame suggesting the presence of a truncated APC protein. We have developed a series of anti-APC antibodies directed against the amino terminal portion of APC. These antibodies have identified a truncated form of APC protein in SW480 cells carrying mutated APC alleles. We will examine a series of tumor cell lines and normal cells for altered forms of APC protein associated with the presence of colorectal cancer. We will concentrate on cell lines in which the structure of APC is known, either wild type of mutated. We will also begin generating antibodies to carboxyl terminal domains in APC that will be used to discriminate full length versus truncated APC proteins. In Phase II, the combination of anti-amino terminal and anti-carboxyl terminal APC antibodies will be used to develop an assay for determination of APC status in clinical samples as a prognostic test for FAP and colorectal cancer.

APC 基因的突变似乎对于腺瘤的形成至关重要 遗传性家族性腺瘤性息肉病 (FAP) 和早期阶段 结直肠癌。 APC 基因分析发现突变 FAP 患者的种系 DNA 中。 超过90%的突变中断 阅读框表明存在截短的 APC 蛋白。 我们开发了一系列针对 APC 的抗体 APC 的氨基末端部分。 这些抗体已鉴定出 携带突变 APC 的 SW480 细胞中 APC 蛋白的截短形式 等位基因。 我们将检查一系列肿瘤细胞系和正常细胞 对于与存在相关的 APC 蛋白的改变形式 结直肠癌。 我们将专注于细胞系，其中 APC的结构是已知的，无论是野生型还是突变型。 我们还将 开始在 APC 中生成针对羧基末端结构域的抗体，该抗体将 可用于区分全长 APC 蛋白与截短的 APC 蛋白。 在 第二阶段，反氨基端与反羧基的结合 末端 APC 抗体将用于开发一种检测方法 确定临床样本中的 APC 状态作为预后测试 FAP 和结直肠癌。