CONTROL OF C-MYC TRANSCRIPTION IN AGGRESSIVE LYMPHOMAS

侵袭性淋巴瘤中 C-MYC 转录的控制

• 批准号: 6102161
• 负责人: LINDA M BOXER
• 金额: $ 21.01万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-29 至 2000-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6102161
• 关键词: B lymphocyte; Burkitt's lymphoma; DNA footprinting; clinical research; crosslink; gene mutation; genetic regulation; genetic transcription; human subject; immunoprecipitation; lymphoma; methylation; molecular oncology; neoplasm /cancer genetics; neoplastic transformation; nucleic acid hybridization; protooncogene; tissue /cell culture; transcription factor; transfection; ultraviolet radiation

I propose to study the mechanism of activation of c-myc at a molecular level in both Burkitt's lymphoma and transformed lymphomas with the t(8;14) translocation. The goal is to reach a better understanding of the mechanisms of malignant transformation of B cells. A. Identification of the molecular mechanisms of transcriptional control of th c-myc gene in B cells. In vivo footprinting and in vitro protein-DNA binding studies will be used to locate regulatory regions important for c-myc expression in normal B cells and in B cell lines that do not contain the t(8;14) translocation. The identity of the proteins that bind in vivo will be determined by a technique of UV crosslinking and immunoprecipitation followed by hybridization analysis. Functional assessment of the sites will be performed with transient transfection experiments in B cell lines. B. Identification of the molecular mechanisms of transcriptional control of the c-myc gene in Burkitt's lymphomas. Conditions have been optimized for the separation of the translocated from the normal c-myc gene in several Burkitt's cell lines. Differences in the binding of transcription factors between the two alleles have been found and these studies will be continued. The identification and characterization of the proteins that bind in vivo will be determined as described in A. The methylation status of each allele will be compared. C. Identification of the molecular mechanisms of transcriptional control of the c-myc and bcl-2 genes in transformed lymphomas and their low grade counterparts. The deregulation of both c-myc and bcl-2 by the immunoglobulin locus will be studied as outlined in B. D. Effect on c-myc expression of mutations identified in the regulatory regions. The effects of any mutations ont he binding of regulatory proteins will be investigated by in vivo and in vitro studies. E. Role of the immunoglobulin locus in the activation of c-myc expression. Model constructs which reproduce the c-myc translocations will be constructed and used to define the important regions of the immunoglobulin locus. The successful completion of this proposal will represent the first instance where the molecular mechanisms involved in the transcriptional activation of an oncogene by translocation are known. The transcription factors that are required for the activation of c-myc will be identified and the regions of the immunoglobulin locus which are responsible for this will be defined.

我建议研究分子上C-MYC激活的机理 伯基特淋巴瘤和T淋巴瘤的水平（8; 14） 易位。 目标是更好地理解 B细胞恶性转化的机制。 A.鉴定转录控制的分子机制 B细胞中的Th c-Myc基因。 将使用体内足迹和体外蛋白-DNA结合研究 要找到对正常B中C-MYC表达很重要的调节区域 细胞和B细胞系中不包含t（8; 14）易位的细胞系。 结合体内结合的蛋白质的身份将由a确定 紫外线交联和免疫沉淀的技术，然后 杂交分析。 站点的功能评估将是 用B细胞系中的瞬时转染实验进行。 B.鉴定转录控制的分子机制 Burkitt淋巴瘤中的C-Myc基因。 条件已优化 为了从正常C-MYC基因中分离 伯基特的几个细胞系。 转录结合的差异 已经发现了两个等位基因之间的因素，这些研究将是 持续。 蛋白质的识别和表征 结合体内将确定如A中所述。甲基化状态 将比较每个等位基因。 C.鉴定转录控制的分子机制 转化的淋巴瘤中的C-Myc和Bcl-2基因及其低等级 同行。 通过 免疫球蛋白基因座将如B中概述。 D.对调节性突变的C-MYC表达的影响 地区。 任何突变的效果都具有调节性的结合 蛋白质将通过体内和体外研究研究。 E.免疫球蛋白基因座在C-MYC表达激活中的作用。 重现C-MYC易位的模型结构将是 构建并用于定义免疫球蛋白的重要区域 轨迹。 该提案的成功完成将代表第一个 实例分子机制涉及转录 已知通过易位激活致癌基因。 转录 将确定激活C-MYC所需的因素 以及负责此的免疫球蛋白基因座的区域 将定义。