IDENTIFICATION OF TRANSFORMING GENES IN MELANOMA

黑色素瘤中转化基因的鉴定

• 批准号: 3202463
• 负责人: TIMOTHY Peter FLEMING
• 金额: $ 21.1万
• 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 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3202463
• 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人患上恶性黑素瘤， 占美国所有新发癌症病例的3%， 这种恶性肿瘤。在过去的十年中，黑色素瘤的发病率 比肺癌以外的任何其他癌症都快， 妇女在美国，个人的终身风险 恶性黑色素瘤在1987年为1：135， 在2000年发病率上升的原因尚不清楚 这是可以理解的，但可能与增加阳光照射有关， 到达表面的紫外线照射量，以及 黑素瘤。 我们实验室最近的努力是为了构建 和cDNA表达系统的实施，该系统已经成功地在 分离在人类恶性肿瘤中表达的激活的癌基因。 人黑色素瘤cDNA文库在NIH/3 T3细胞中的表达克隆 鉴定了一种新的转化基因，称为tim-1，用于转化 黑素瘤该基因与其他基因没有整体序列同源性。 Genebank中有一个102 bp的区域， 与位于基因db 1，bcr，CDC 24，ect-2， CDC 25和VAV。这一同源区域代表了一个扩展的 这些基因家族可能对生长控制很重要。 探讨该基因在恶性肿瘤发病机制中的作用， 黑色素瘤，我们提出以下具体目标： #1.我们将转染tim-1基因的cDNA导入永生化小鼠， 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 表情还将检测正常人中Tim-1的表达 组织和发育中的小鼠。 #3.将分离全长tim-1 cDNA（6.8 kb）， 转化活性，相对于克隆的转化cDNA（2.4kb）， 将被审查。 #4.定义tim-1基因的染色体定位并表征 TIM-1蛋白。