INTESTINAL CELL GROWTH CONTROL--ROLE OF TYROSINE KINASE

肠细胞生长控制——酪氨酸激酶的作用

• 批准号: 3464440
• 负责人: CHRISTINE Ann CARTWRIGHT
• 金额: $ 10.89万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-06-15 至 1996-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3464440
• 关键词: affinity chromatography; antisense nucleic acid; carcinogenesis; cell growth regulation; chickens; colon neoplasms; enzyme mechanism; fibroblasts; gastrointestinal epithelium; gene mutation; human subject; intestines; oncogenes; phosphorylation; polymerase chain reaction; posttranslational modifications; preneoplastic state; protein tyrosine kinase; ulcerative colitis; western blottings

Regulation of intestinal cell growth and the loss of that regulation during malignant transformation of cells involves a complex series of molecular events. Our objective is to determine the role of pp60c-scr and related protein-tyrosine kinases in this regulatory process. Historically, a strong correlation exists between elevated pp60src kinase activity and cell transformation. We determined that pp60c-src isolated from human colon carcinomas, malignant colonic polyps, and benign polyps at greatest risk for developing cancer, has higher specific activity than pp60c-src from normal mucosa. Our results also show that pp60c-src activity decreases as primary cells differentiate along the crypt-villus axis of chicken intestine. Together, the data suggest that down-regulation of c-src is important for differentiation, and deregulation of c-src is important for transformation of intestinal epithelia. We will study mechanisms that regulate pp60c-src activity in intestinal cells. We, and others, have identified three mechanisms that regulate pp60c-src activity in other cells: phosphorylation of pp60c-src, association of pp60c-src with another protein, or mutation within the coding region of c-src. Posttranslational events that are known to alter pp60c-src activity will be studied in cells along the crypt-villus axis of chicken intestine. c-src will be isolated from colon carcinomas which have elevated pp60c-src activity, and its coding region will be analyzed for somatic mutations and tested for oncogenic potential in fibroblasts. pp60c-src activity will be measured in colonic mucosa of patients with ulcerative colitis which, like adenomatous polyps, has malignant potential. Finally, the activity of pp60c-src -related tyrosine kinases will be measured and the substrates of tyrosine kinases will be identified in colon carcinoma cells. Our aim is to understand the role of pp60c-src and related kinases in regulating growth of intestinal cells. By addressing basic mechanisms of growth control in normal cells, we can elucidate pathogenic mechanisms in human disease, namely, colonic carcinogenesis. We hope to identify early events that perturb normal regulatory pathways in the precancerous mucosa of colonic polyps and ulcerative colitis. The results will impact on prevention, diagnosis and treatment of colon cancer.

调节肠细胞生长和该调节的损失 在细胞的恶性转化过程中，涉及一系列复杂的一系列 分子事件。 我们的目标是确定PP60C-SCR的作用 和相关的蛋白质酪氨酸激酶在此调节过程中。 从历史上看，升高的PP60SRC之间存在很强的相关性 激酶活性和细胞转化。 我们确定PP60C-SRC 从人类结肠癌，恶性结肠息肉和 良性的息肉有最大的癌症风险，具有更高的 比正常粘膜的PP60C-SRC比特定活性。 我们的结果也是如此 表明PP60C-SRC活性随着原代细胞的分化而降低 沿着鸡肉肠道的地下室villus轴。 在一起，数据 表明C-SRC的下调对于分化很重要， C-SRC的放松管制对于肠道转化很重要 上皮。 我们将研究调节肠道PP60C-SRC活性的机制 细胞。 我们和其他人已经确定了调节的三种机制 其他细胞中的PP60C-SRC活性：PP60C-SRC的磷酸化， PP60C-SRC与另一种蛋白质的关联，或 C-SRC的编码区域。 已知的翻译后事件 Alter PP60C-SRC活性将在沿着隐窝村的细胞中进行研究 鸡肠轴。 C-SRC将从结肠癌中分离出来 PP60C-SRC活性升高，其编码区域将是 分析了体细胞突变并测试了致癌潜力 成纤维细胞。 PP60C-SRC活性将在结肠粘膜中测量 溃疡性结肠炎患者，像腺瘤息肉一样 恶性潜力。 最后，PP60C -SRC相关的活性 将测量酪氨酸激酶并测量酪氨酸激酶的底物 将在结肠癌细胞中鉴定。 我们的目的是了解PP60C-SRC和相关激酶在 调节肠细胞的生长。 通过解决基本机制 正常细胞的生长控制，我们可以阐明致病性 人类疾病的机制，即结肠致癌。 我们希望 确定扰动正常调节途径的早期事件 结肠息肉和溃疡性结肠炎的癌性粘膜。 这 结果将影响结肠的预防，诊断和治疗 癌症。