MOLECULAR/CYTOGENETICS ESTROGEN-INDUCED RENAL NEOPLASIA

我们的总体目标是阐明早期的细胞遗传学和分子生物学 遗传事件强调雌激素诱导的肾肿瘤发生在 仓鼠肾，并将这些发现与仓鼠肾肿瘤进行比较， 我们的两个“半死不活”的雌激素衍生的肾细胞系。 的 该应用的优势在于两口井的经验 建立了与仓鼠系统合作的小组， 能力. 合作小组将开发一套表达方式 标签位点（ETS）建立仓鼠基因组的物理图谱， 最后一张基因图谱将被用来识别受影响的人 在肾肿瘤发生过程中基因发生改变。 为了实现这些目标， 提出以下研究建议：1.系统性细胞遗传学分析 肾肿瘤和肾在雌激素诱导的肿瘤发生。 这 将通过对各种时间关系的详细研究来完成 肾肿瘤发生过程中的细胞遗传学改变。 非随机染色体 变更（收益和损失）将在每月的时间间隔确定， 雌激素治疗，它们将与非随机的数值变化有关， 脆弱地点的频率和位置将根据 仓鼠肾染色体雌激素化肾脏，肾肿瘤， DES衍生的肾细胞系。2.筛选cDNA探针 原癌基因和其他相关受体蛋白表达 雌激素诱导的肿瘤发生。 这将通过杂交来完成， 从仓鼠肾脏在不同阶段的新合成的RNA 雌激素处理的cDNA克隆阵列。 克隆人会 将通过北方杂交来证实表现出改变的表达。 这些研究将补充具体目标1和4。研究 cDNA探针的细胞表达将是原位杂交的前奏 在染色体上和雌激素化的 肾脏和肿瘤细胞。3.创建一组带有表情标签的网站 (ETS)建立叙利亚仓鼠基因组的物理图谱。 仓鼠 将用来自转录单位随机样本的探针进行映射。 一个仓鼠基因图谱将被建立，然后将被用作道路 以确定致癌过程中涉及的基因。cDNA克隆 将被分离，并且从两端测序150-300 bp区域。 序列信息将用于搜索基因组数据库。4. 用ETS和其他相关位点建立遗传图谱。 发展 同线群 这将通过仓鼠的原位杂交来完成。 中期扩散。 将通过比较生成同线组 将仓鼠数据分析到小鼠和人类基因组数据库。

Our overall aim is to elucidate the early cytogenetic and molecular genetic events underlining estrogen-induced renal tumorigenesis in the hamster kidney and compare these findings to the hamster renal tumor and to our two "semi-immortal" estrogen-derived kidney cell lines. The strength of this application resides in the experience of two well established groups working with hamster systems with complementary abilities. The collaborating group will develop a battery of expression tagged sites (ETS) to established a physical map of the hamster genome, and eventually a genetic map which will be used to identify the affected genes being altered during renal tumorigenesis. To pursued these aims, the following studies are proposed: 1. Systematic cytogenetic analyses of the kidney tumor and kidney during estrogen-induced tumorigenesis. This will be done by detail studies of the temporal relationship of various cytogenetic alterations during renal tumorigenesis. Nonrandom chromosome alterations (gains and losses) will be determined at monthly intervals of estrogen treatment, they will be related to nonrandom numerical changes, and the frequency and location of fragile sites will be determined on hamster kidney chromosomes in estrogenized kidneys, renal tumor, and DES-derived kidney cell lines. 2. Screen for cDNA probes for protooncogene and other pertinent receptor protein expression during estrogen-induced tumorigenesis. This will be done by hybridization of newly synthesized RNA from hamster kidneys at different stages of estrogen treatment to an array of cDNA clones. The clones that will exhibit changed expression will be confirmed by Northern hybridization. These studies will complement Specific Aims 1 and 4. The study of cellular expression of the cDNA probes will be a prelude to in-situ hybridization studies both on the chromosome and within estrogenized kidney and tumor cells. 3. Create a battery of expression tagged sites (ETS) to establish a physical map of the Syrian hamster genome. Hamsters will be mapped with probes from a random sample of transcription units. A hamster genetic map will be established and will then be used as a road map to identify genes involved in the carcinogenic process. cDNA clones will be isolated, and 150-300 bp regions sequenced from both ends. Sequence information will be used to search the genome database. 4. Establish a genetic map with ETS and other relevant loci. Develop syntenic groups. This will be done by in situ hybridization of hamster metaphase spreads. Syntenic groups will be generated by comparison analysis of hamster data to the mouse and human genome database.