GROWTH SUPPRESSOR GENES IN METASTATIC MELANOMA

转移性黑色素瘤中的生长抑制基因

• 批准号: 3423628
• 负责人: FREDERICK T BOYD
• 金额: $ 5.17万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-08-01 至 1993-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3423628
• 关键词: cell adhesion; cell growth regulation; cellular oncology; complementary DNA; extracellular matrix; gene expression; genetic library; growth inhibitors; melanoma; metastasis; molecular cloning; neoplasm /cancer genetics; neoplasm /cancer invasiveness; neoplastic cell; phenotype; plasmids; polymerase chain reaction; transfection; tumor suppressor genes

Our ultimate goal is to identify genes which are active in promoting metastatic behavior of tumor cells. We propose to use two recently developed phenotypic selection ethods to identify cDNAs which are expressed in metastatic tumor cells and which may be involved in particular aspects of the metastatic process. We will apply our recently developed assay for the positive selection of slowly growing cells, coupled with classical gene transfer and differential expression methodology, to identify growth suppressor genes which are differentially expressed in metastatic versus non-metastatic melanoma cells. In addition will attempt to adapt a method for selecting cell variants which are capable of invasion as election method for genes which are involved in the invasion process. The advantage to is approach is that interesting genes are initially selected on the basis of a desired phenotype, not merely differential expression. 1. Determine the effect of our previously isolated growth inhibitory plasmids on the growth characteristics and metastatic behavior of established high and low metastatic potential melanoma lines. 2. Identify genes which are expressed in metastatic tumor cells and non-metastatic tumor cells which are capable of inhibiting cell proliferation. 3. Develop a phenotypic selection protocol for the isolation of invasion promoting genes. 4. Assess the patterns of expression of putative growth inhibitory or invasion promoting genes to determine whether they are differentially expressed in metastatic cells versus non-metastatic cells. 5. Determine if the expression of any putative growth inhibitory or invasion promoting genes modulates components of the extracellular matrix and matrix receptors which have been previously correlated with the metastatic phenotype.

我们的最终目标是识别积极促进 肿瘤细胞的转移行为。我们建议最近使用两个 开发了表型选择方法来鉴定 cDNA 在转移性肿瘤细胞中表达，可能参与 转移过程的特定方面。 我们将应用我们的 最近开发的用于缓慢生长的阳性选择的测定法 细胞，加上经典的基因转移和差异表达 方法，以确定生长抑制基因 在转移性黑色素瘤与非转移性黑色素瘤中表达差异 细胞。 此外将尝试采用一种选择细胞的方法 能够作为基因选择方法入侵的变体 都参与了入侵过程。 的优点是 方法是最初选择感兴趣的基因 所需的表型，而不仅仅是差异表达。 1. 确定我们之前分离的生长抑制剂的效果 质粒对生长特性和转移行为的影响 建立了高和低转移潜能黑色素瘤系。 2. 鉴定在转移性肿瘤细胞中表达的基因 能够抑制细胞的非转移性肿瘤细胞 增殖。 3. 制定用于分离入侵的表型选择方案 促进基因。 4. 评估假定的生长抑制或生长抑制的表达模式 入侵促进基因以确定它们是否存在差异 在转移细胞与非转移细胞中表达。 5. 确定是否存在任何假定的生长抑制或 侵袭促进基因调节细胞外成分 先前已与相关的基质和基质受体 转移表型。