MECHANISM OF ONCOGENE ACTIVATION IN MOUSE LYMPHOMAS

小鼠淋巴瘤中癌基因激活的机制

• 批准号: 3180627
• 负责人: ELIZABETH W. NEWCOMB
• 金额: $ 24.38万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-07-01 至 1995-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3180627
• 关键词: athymic mouse; chromosome translocation; endonuclease; gamma radiation; gene expression; genetic markers; genetic promoter element; lymphoma; molecular cloning; neoplasm /cancer genetics; northern blottings; oncogenes; polymerase chain reaction; radiation carcinogenesis; radiation genetics; radiation related neoplasm /cancer; restriction fragment length polymorphism; restriction mapping; transcription factor; transfection; transposon /insertion element; western blottings

In this application we will continue to study the transforming genes associated with gamma irradiation induced thymic lymphomas in C57BL/6J mice. We have reported a specific marker chromosome consisting of a translocation between chromosomes one and five was observed in 5 of 12 gamma irradiation induced thymic lymphomas. DNA from these 5 thymic lymphomas contains a novel non-ras transforming activity detected by DNA-mediated-gene-transfer using a sensitive tumorigenicity assay in nude mice to score for the presence of transforming DNA sequences. We will clone the novel transforming gene(s), using the nude mouse tumorigenicity assay to monitor the biological activity of the cloned DNA fragments. A detailed restriction endonuclease map of the oncogene(s) will be constructed. A subclone of the biologicallly active DNA fragment, consisting of single copy mouse DNA, will be used in Southern and Northern blot analysis to determine the number and arrangement of the gene(s) in the mouse genome and whether the gene(s) is expressed in mRNA. To determine the mechanism of oncogene activation in the 5 primary radiation-induced thymic lymphomas containing the marker chromosome, the five oncogenes, or relevant portions of the transforming DNA sequences, will be cloned. If the 5 genes are the same, fine detail analysis will determine the nature of the chromosomal change(s) resulting in oncogene activation. If the 5 genes are different, then cloning and DNA sequence analysis will determine the different mechanisms of oncogene activation. Finally, we will assess the timing of oncogene activation in mouse thymocytes at intervals following gamma irradiation treatment. If the activated oncogene is associated with the presence of the marker chromosome, then the polymerase chain reaction (PCR) technique will be used to detect the breakpoint of the chromosome translocation at different stages of disease. Experimental animal models have been extremely useful in gaining insights into some of the steps involved in disease progression. This study should elucidate some of the mechanisms involved in oncogene activation as they are related to the translocation of chromosomal segments, and has the potential for identifying a new oncogene(s) involved in lymphoid malignancies.

在此应用中，我们将继续研究转化基因 与γ辐照在C57BL/6J中诱导的胸腺淋巴瘤有关 老鼠。我们报告了一个特定的标记染色体，该染色体由 在12中的5个中观察到染色体1和5之间的易位 γ辐照诱导的胸腺淋巴瘤。这5个百里香的DNA 淋巴瘤包含一种新的非RAS转化活性，该活性检测到 DNA介导的基因转移，使用裸体中敏感的肿瘤性测定法 小鼠得分在转化的DNA序列的存在下得分。我们会克隆 新颖的转化基因，使用裸小鼠肿瘤性测定法 监测克隆的DNA片段的生物学活性。详细的 将构建癌基因的限制性核酸内切酶图。一个 生物活性DNA片段的亚克隆，由单个 复制鼠标DNA，将在南部和北印迹分析中使用 确定小鼠基因组中基因的数量和排列 是否在mRNA中表达基因。确定机制 5个原代辐射诱导的胸腺淋巴瘤中的癌基因激活 包含标记染色体，五个癌基因或相关部分 转化的DNA序列将被克隆。如果5个基因是 同样，细节分析将确定染色体的性质 变化导致致癌基因激活。如果5个基因不同， 然后克隆和DNA序列分析将确定不同的 癌基活化的机制。最后，我们将评估 伽马后，小鼠胸腺细胞中的癌基因激活 辐照处理。如果激活的癌基因与 标记染色体的存在，然后是聚合酶链反应（PCR） 技术将用于检测染色体的断点 疾病不同阶段的易位。实验动物模型 在获得一些步骤方面非常有用 参与疾病进展。这项研究应阐明一些 与癌基激活有关的机制，因为它们与 染色体片段的易位，并具有 确定涉及淋巴恶性肿瘤的新癌基因。