Neoplastic transformation of melanocytes by Grm1

Grm1 对黑素细胞的肿瘤转化

• 批准号: 6916441
• 负责人: Suzie Chen
• 金额: $ 24.99万
• 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/6916441
• 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由g蛋白偶联的7跨膜结构域受体Grm1异位表达引起的黑素细胞转化的分子机制，并检查在这些细胞中继续表达Grm1介导其致癌潜能的必要性。我们的目标是基于我们最初的假设和我们最近发表的假设的证实。此前，我们提出使用具有自发性黑色素瘤发展易感性的转基因小鼠TG-3作为模型系统来研究这种疾病。我们确定了由于转基因整合而改变的宿主序列为Grm1。我们发现Grm1在肿瘤中异常表达，而在对照样本中没有。我们假设Grm1与TG- 3中黑色素瘤的发生有关。然后，我们通过将Grm1的cDNA置于黑素细胞特异性启动子多巴胺自变酶（dopachrome tautomerase, Dct）的调控下，产生了一种新的转基因系，表达了仅针对黑素细胞的Grm1l，从而验证了这一假设。这个新的Dct-Grm1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，我们还证实GRM1在一些人类黑色素瘤细胞系和活检样本中表达，但在正常黑色素细胞或良性痣中不表达。因此，我们已经明确地证明了Grm1异常表达和黑色素瘤在我们的系统中发展的病因学作用。基于这些结果，我们建议开始一组实验，使用来自黑素细胞肿瘤的细胞，开始使用我们的模型系统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。目的：研究和表征Grm1l在维持转化表型中持续表达的要求。2. 扩展并验证Grm1在人类黑色素瘤中的致癌作用。3. 探讨Grm1在TG-3小鼠肿瘤发生和发展过程中的持续表达。4. 探讨MAPK信号在小鼠模型系统中的作用。