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。 雌激素诱导大鼠肾脏肿瘤发生的遗传学事件 并将这些发现与仓鼠肾脏肿瘤和 我们的两个“半永生”雌激素来源的肾脏细胞系。这个 这个应用的优势在于两口井的体验 与仓鼠系统合作的现有小组具有互补性 超能力。合作小组将开发一系列表达 标记位点(ETS)来建立仓鼠基因组的物理图谱， 最终得到一张基因图谱，用来识别受影响的人 肾脏肿瘤发生过程中基因的改变。为了追求这些目标， 主要研究内容如下：1.系统的细胞遗传学分析 雌激素诱导肿瘤发生过程中肾肿瘤与肾脏的关系。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(损益)将按月确定 雌激素治疗，它们将与非随机的数字变化有关， 脆弱部位的频率和位置将在 地鼠肾染色体在雌激素肾、肾肿瘤和 DES来源的肾脏细胞系。2.筛选cDNAs探针 原癌基因和其他相关受体蛋白的表达 雌激素诱导的肿瘤发生。这将通过杂交的方式完成 新合成的地鼠肾脏不同阶段的RNA 雌激素对一系列cdna克隆的处理。克隆人将 表现出的表达变化将通过Northern杂交得到证实。 这些研究将补充具体目标1和4。 CDNAs探针的细胞表达将是原位研究的前奏 染色体和雌激素化的杂交研究 肾脏和肿瘤细胞。3.创建一系列带有表情标签的站点 (ETS)建立叙利亚仓鼠基因组的物理图谱。仓鼠 将与转录单位的随机样本中的探针进行映射。 仓鼠基因图谱将被建立，然后将被用作道路 绘制图谱以识别参与致癌过程的基因。CDNA克隆 将被分离，并从两端对150-300个碱基区进行测序。 序列信息将用于搜索基因组数据库。4. 建立与ETS等相关基因座的遗传图谱。发展 同音群。这将通过仓鼠的原位杂交来完成 中期展开。将通过比较生成合成基团 分析仓鼠的数据，以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.