IDENTIFICATION OF TRANSFORMING GENES IN MELANOMA

黑色素瘤中转化基因的鉴定

• 批准号: 2099006
• 负责人: TIMOTHY Peter FLEMING
• 金额: $ 21.11万
• 依托单位: WASHINGTON UNIVERSITY
• 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 1998-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2099006
• 关键词: 3T3 cells; athymic mouse; cell line; chromosomes; complementary DNA; developmental genetics; embryo /fetus; gene expression; genetic library; genetic mapping; human tissue; immunoprecipitation; laboratory rabbit; melanocyte; melanoma; metastasis; molecular cloning; neoplastic transformation; northern blottings; nucleic acid sequence; oncogenes; oncoproteins; protein structure; tissue /cell culture; transfection

In 1990 nearly 30,000 individuals developed malignant melanoma, representing 3% of all new cancer cases in the U.S. and 6000 people died of this malignancy. In the past decade, the incidence of melanoma increased faster than that of any other cancer except lung cancer in women. The lifetime risks of an individual in the U.S. developing malignant melanoma was 1:135 in 1987 and is projected to be 1:90 by the year 2000. The reasons for the increased incidence are not clearly understood but likely relate to increased exposure to sunlight, increased amount of Uv irradiation reaching the surface, and earlier detection of melanomas. Recent efforts in our laboratory have been directed to the construction and implementation of a cDNA expression system that has been successful in the isolation of activated oncogenes expressed in human malignancies. Expression cloning of a human melanoma cDNA library into NIH/3T3 cells has identified a novel transforming gene, termed tim-1, for transforming in melanoma. This gene has no overall sequence homology to any genes in Genebank, however there is a 102 bp region that is approximately 30% homologous to a common site located on the genes dbl, bcr, CDC 24, ect-2, CDC 25, and vav. This region of homology is representive of an expanding family of genes which may be important to growth control. To investigate the role this gene has in the pathogenesis of malignant melanoma, we propose the following specific aims: #1. We will transfect the transforming tim-1 cDNA into immortalized mouse melanocytes and determine its effect on tumorigenicity #2. To investigate the role the tim-1 gene may have clinically, we will screen melanoma specimens, metastases, and cell lines for tim-1 gene expression. The expression of tim-1 will also be examined in normal human tissue and in the developing mouse. #3. The full-length tim-1 cDNA (6.8 kb) will be isolated and its transforming activity, relative to the cloned transforming cDNA (2.4 kb), will be examined. #4. Define the chromosomal localization of the tim-1 gene and characterize the tim-1 protein.

1990 年，近 30,000 人罹患恶性黑色素瘤， 占美国所有新癌症病例的 3%，6000 人死亡 这种恶性肿瘤。近十年来，黑色素瘤的发病率 增长速度快于除肺癌以外的任何其他癌症 女性。在美国，个人一生中面临的风险 1987 年恶性黑色素瘤的发病率为 1:135，预计到 2020 年将达到 1:90 2000年发病率增加的原因尚不清楚 理解，但可能与增加暴露在阳光下、增加 到达表面的紫外线照射量，以及早期检测 黑色素瘤。 我们实验室最近的努力方向是建设 并成功实施了 cDNA 表达系统 分离人类恶性肿瘤中表达的活化癌基因。 将人黑色素瘤 cDNA 文库表达克隆到 NIH/3T3 细胞中 鉴定出一种新的转化基因，称为 tim-1，用于转化 黑色素瘤。该基因与任何基因都没有整体序列同源性 然而 Genebank 有一个 102 bp 的区域，大约占 30% 与位于基因 dbl、bcr、CDC 24、ect-2 上的共同位点同源， CDC 25 和 vav。该同源区域代表了不断扩大的 可能对生长控制很重要的基因家族。 研究该基因在恶性肿瘤发病机制中的作用 针对黑色素瘤，我们提出以下具体目标： #1.我们将转化的tim-1 cDNA转染到永生化小鼠中 黑素细胞并确定其对致瘤性的影响 #2.为了研究 tim-1 基因在临床上可能发挥的作用，我们将 筛选黑色素瘤标本、转移瘤和细胞系中的 tim-1 基因 表达。 Tim-1 的表达也将在正常人中进行检测 组织和发育中的小鼠。 ＃3。将分离出全长 tim-1 cDNA (6.8 kb)，并将其 相对于克隆的转化 cDNA (2.4 kb) 的转化活性， 将接受检查。 ＃4。定义 tim-1 基因的染色体定位并表征 tim-1 蛋白。