DEVELOPMENTAL & NEOPLASTIC GENE EXPRESSION IN THE LIVER

发展型

• 批准号: 6173354
• 负责人: JOSEPH D LOCKER
• 金额: $ 25.44万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-07-01 至 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6173354
• 关键词: alpha fetoprotein; cell transformation; embryo /fetus tissue /cell culture; gene deletion mutation; gene induction /repression; gene targeting; genetic mapping; genetic regulation; genetically modified animals; hepatocellular carcinoma; histogenesis; human tissue; laboratory mouse; laboratory rat; liver cells; molecular cloning; neoplasm /cancer genetics; nucleic acid sequence; plasmids; transcription factor; yeast two hybrid system

DESCRIPTION: The investigators have now cloned the cDNA encoding a homeobox transcription factor related to Drosophila NK2. As the seventh vertebrae factor in this family, the new factor is tentatively designated PCF/Nkx2.7. Nkx 2.1 to 2.6 are all developmental regulators of organogenesis, and studies are proposed to determine the role of PCF/Nkx 2.7 in liver development and neoplasia. Aim 1 studies will focus on the new gene: the transcriptional regulatory controls will be characterized, and in addition, a closely related gene will be cloned and sequenced. In Aim 2, the properties of the PCF/Nkx 2.7 molecule will be analyzed by deletion of peptide domains and GAL4 gene fusion. Subsequently, the binding partners will be determined using direct assays and the yeast two-hybrid system. Aim 3 evaluates the role of PCF/Nk 2.7 in development through analysis of embryos and cell cultures. A mouse inducible gene targeting model will be used to confine gene knock-out to the foregut, the embryonic region that gives rise to the liver, or to the early embryo. Finally, Aim 4 deals with cancer -related aspects. A rearranged PCF/Nkx 2.7 has been found in a human hepatocarcinoma cell line, suggested a possible role in cell transformation. This putative translocation will be sequenced and mapped. Studies will evaluate PCF/Nkx 2.7 as a transforming gene in cell lines, and in primary culture of hepatocytes. Together these Aims represent a comprehensive approach to this new gene, based on the model that abnormal reactivation of PCF/Nkx 2.7 recapitulates developmental controls to transform liver cells.

描述：研究人员现已克隆了编码同源盒的 cDNA 与果蝇 NK2 相关的转录因子。 作为第七椎骨 该因子家族中的新因子暂定为PCF/Nkx2.7。 Nkx 2.1 至 2.6 都是器官发生的发育调节因子，并且 建议进行研究以确定 PCF/Nkx 2.7 在肝脏中的作用 发育和瘤形成。 目标 1 研究将重点关注新基因： 转录调控控制将得到表征，此外， 密切相关的基因将被克隆并测序。 在目标 2 中， PCF/Nkx 2.7 分子的特性将通过删除来分析 肽结构域和 GAL4 基因融合。 随后，绑定伙伴 将使用直接测定和酵母双杂交系统来确定。 目的 3 通过分析评估 PCF/Nk 2.7 在发育中的作用 胚胎和细胞培养物。 小鼠诱导基因打靶模型将是 用于将基因敲除限制在前肠，即胚胎区域 产生肝脏或早期胚胎。 最后，目标 4 涉及 癌症相关方面。 在人类体内发现了重排的 PCF/Nkx 2.7 肝癌细胞系，表明在细胞转化中可能发挥作用。 这种假定的易位将被测序和绘制。 研究将 评估 PCF/Nkx 2.7 作为细胞系和原代细胞中的转化基因 肝细胞培养。 这些目标共同代表了一个全面的 基于异常重新激活的模型来研究这个新基因 PCF/Nkx 2.7 概括了转化肝细胞的发育控制。