MOLECULAR GENETIC STUDIES OF THYROID CELL LINES

甲状腺细胞系的分子遗传学研究

• 批准号: 2143278
• 负责人: DUNCAN C FERGUSON
• 金额: $ 3.15万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-06-01 至 1994-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2143278
• 关键词: cell cycle; cell differentiation; clone cells; gene expression; genetic manipulation; goiter; molecular genetics; northern blottings; nucleic acid hybridization; oncogenes; peroxidases; protooncogene; southern blotting; thyroglobulin; thyroid disorder; thyroid gland; transcription factor; tumor suppressor genes

The longterm objectives of these studies are to establish spontaneous feline hyperfunctional adenomatous goiter (toxic nodular goiter) as a model for human benign thyroid tumors, and to probe for molecular genetic abnormalities associated with the changeover from normal to adenomatous growth with maintained differentiated function. The proposed studies focus upon the factors controlling the normal cell cycle of the feline adenomatous thyrocyte cell lines. Particular attention will be paid to possible alterations in oncogene and anti-oncogene structure and expression. In order to acquire the necessary techniques to accomplish this longterm goal, this proposal outlines pilot work which will allow the principal investigator to incorporate molecular genetic techniques into ongoing and future research projects in thyroidology. In experiments involving the feline adenomatous thyrocyte cell lines PETCAT-l,-2,-3,-4, and ROMCAT-1, this proposal has the following specific aims: 1. For the visiting researcher to become familiar with the molecular genetic techniques of RNA and DNA extraction and purification, nucleic acid electrophoresis, dot blot assays, Northern and Southern blotting and hybridization techniques, and to learn about application of molecular genetic techniques to the study of cell cycle events. 2. To evaluate alterations in DNA structure and basal mRNA expression of known proto-oncogenes, transcription factors, tumor suppression factors, and signal transduction factors. 3. To study mRNA expression kinetics of growth-related genetic factors and compare them with the expression of factors associated with thyrocyte differentiation under conditions stimulatory for growth.

这些研究的长期目标是建立自发的 猫功能亢进性腺瘤性甲状腺肿（毒性结节性甲状腺肿） 人类良性甲状腺肿瘤模型，并探讨分子遗传学 与从正常转变为腺瘤相关的异常 生长并保持分化功能。 拟议的研究 关注控制猫科动物正常细胞周期的因素 腺瘤甲状腺细胞系。 将特别关注 癌基因和抗癌基因结构可能发生改变， 表达。 为了获得完成任务所需的技术 为了实现这一长期目标，该提案概述了试点工作，这将使 结合分子遗传学技术的主要研究者 正在进行和未来的甲状腺学研究项目。 在涉及猫腺瘤甲状腺细胞系的实验中 PETCAT-l、-2、-3、-4和ROMCAT-1，本提案有以下具体内容 目标： 1. 供客座研究员熟悉分子 RNA和DNA提取和纯化、核酸遗传技术 酸性电泳、斑点印迹分析、Northern 和 Southern 印迹以及 杂交技术，并了解分子杂交技术的应用 遗传技术用于研究细胞周期事件。 2. 评估 DNA 结构和基础 mRNA 表达的变化 已知的原癌基因、转录因子、肿瘤抑制因子、 和信号转导因子。 3. 研究生长相关遗传因子的mRNA表达动力学 并与甲状腺细胞相关因子的表达进行比较 在刺激生长的条件下分化。