Neoplastic transformation of melanocytes by Grm1

Grm1 对黑素细胞的肿瘤转化

• 批准号: 7071111
• 负责人: Suzie Chen
• 金额: $ 24.39万
• 依托单位: RUTGERS, THE STATE UNIV OF N.J.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7071111
• 关键词: RNA interference; apoptosis; athymic mouse; biological signal transduction; biopsy; cell line; cell surface receptors; cellular oncology; clinical research; disease /disorder model; genetically modified animals; human tissue; laboratory mouse; melanocyte; melanoma; mitogen activated protein kinase; molecular oncology; neoplasm /cancer genetics; neoplastic process; neoplastic transformation; oncogenes; receptor expression; tissue /cell culture; transfection

DESCRIPTION (provided by applicant): The Aims of this proposal are to study the molecular mechanisms of transformation of melanocytic cells elicited by the ectopic expression of the G-protein-coupled seven transmembrane-domain receptor, Grm1, and to examine the requirement of continuing expression of Grm1in mediating its oncogenic potential in these cells. Our Aims are based on our initial hypothesis and confirmation of the hypothesis in our recent publication. Previously we proposed to use a line of transgenic mice, TG-3, with a predisposition to spontaneous melanoma development as a model system to study this disease. We identified the altered host sequences due to the integration of the transgene to be Grm1. We showed the aberrant expression of Grm1 in tumor but not control samples. We hypothesized that Grm1 is responsible for the genesis of melanoma in TG- 3. We then tested this hypothesis by generating a new transgenic line with expression of Grm1l targeted only to melanocytes, by placing the cDNA of Grm1 under the regulation of the melanocyte specific promoter, dopachrome tautomerase (Dct). This new Dct-Grm1 transgenic line (the E line) showed predisposition to melanoma development similar to that of TG-3. Furthermore, we also demonstrated the expression of GRM1 in some human melanoma cell lines and biopsy samples but not in normal melanocytes or benign nevi. Thus, we have demonstrated unequivocally the etiological role of aberrant Grm1 expression and melanoma development in our system. Based on these results, we propose to initial a set of experiments using cells derived from melanocytic tumors to begin to unravel the complicated yet well coordinated network of cellular transformation using our model system. Our specific aims are: 1. To investigate and characterize the requirement for continuing expression of Grm1l in the maintenance of the transformed phenotypes. 2. To extend and validate the oncogenic role of Grm1 in human melanoma. 3. To examine the requirement for continuous Grm1 expression in onset and progression of tumor in TG-3 mice. 4. To explore MAPK signaling in our mouse model system.

描述（由申请人提供）：本提案的目的是研究G蛋白偶联7跨膜结构域受体Grm 1的异位表达引起的黑素细胞转化的分子机制，并检查Grm 1在这些细胞中介导其致癌潜力的持续表达的必要性。我们的目标是基于我们最初的假设和我们最近出版的假设的确认。以前，我们建议使用一个线的转基因小鼠，TG-3，具有自发性黑色素瘤发展的倾向作为模型系统来研究这种疾病。我们鉴定了由于转基因的整合而改变的宿主序列为Grm 1。我们发现Grm 1在肿瘤中异常表达，而在对照样品中没有。我们假设Grm 1负责TG- 3中黑色素瘤的发生。然后，我们通过将Grm 1的cDNA置于黑素细胞特异性启动子多巴色素互变异构酶（Dct）的调控下，产生一种新的表达Grm 11的转基因株系，来测试这一假设。这种新的Dct-Grm 1转基因系（E系）显示出与TG-3相似的黑色素瘤发展倾向。此外，我们还证明了GRM 1在一些人黑色素瘤细胞系和活检样本中的表达，但在正常黑素细胞或良性痣中不表达。因此，我们已经明确地证明了Grm 1表达异常和黑色素瘤的发展在我们的系统中的病因作用。基于这些结果，我们建议使用来自黑素细胞肿瘤的细胞启动一组实验，以开始使用我们的模型系统来解开复杂但协调良好的细胞转化网络。我们的具体目标是：1.研究和表征在转化表型的维持中持续表达Grm 1 l的需要。2.扩展并验证Grm 1在人黑色素瘤中的致癌作用。3.研究TG-3小鼠肿瘤发生和发展对Grm 1持续表达的需求。4.探讨MAPK信号在小鼠模型系统中的作用。