NEONATAL DES INDUCED UTERINE DYSPLASIA/NEOPLASIA

新生儿 DES 诱发的子宫发育不良/肿瘤

• 批准号: 2100950
• 负责人: WILLIAM J HENDRY
• 金额: $ 15.11万
• 依托单位: WICHITA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-10 至 1996-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2100950
• 关键词: cheek pouch technique; diethylstilbestrol; estrogens; hamsters; histogenesis; hormone regulation /control mechanism; immunocytochemistry; immunoprecipitation; in situ hybridization; neoplastic growth; northern blottings; ovariectomy; protooncogene; regulatory gene; reproductive development; tissue /cell culture; uterus neoplasms; western blottings

When hamsters are healed neonatally with the synthetic estrogen, dethylstibestrol (UES), their uteri consistent exhibit a severe hyperplastic/neoplastic response to estrogen in adulthood. One of two alternatives working hypotheses should explain this phenomenon: 1) Direct Action The cellular physiology and/or composition of the neonatal hamster uterus is directly and permanently altered by the DES insult such that the adult organism overall proliferative response to estrogen becomes atypical, or 2) Indirect Action Uterotrophic activity is mediated by estrogen primarily through or in conjunction with other unidentified factors, and neonatal DES treatment permanently alters the level or functional activity of such factors. Testing these hypotheses will begin (Specific Aim #1) by monitoring the morphogenesis of neonatal uteri from control and DES healed donot animals that are transplanted into the contralateral cheek pouches of control and DES-treated mature hosts that are ovariectomized and estrogen- replaced. To confirm and extend the findings (Specific Aim #2), homotypic and heterotypic recombination of uterine stroma and epithelium from control and DES-treated animals will be performed and their morphogenesis will be studied: a) in vitro using media supplemented with uterine tissue extracts and/or serum from the same host groups used in Specific Aim #1 and b) in vivo (within the cheek pouch) using the same host groups as in Specific Aim #1. Because this combination of approaches relies on few, if any, a prior assumptions and accommodates both in vivo and in vitro observations, firm conclusions should be reached about which alternative hypothesis is most valid. Lastly (Specific Aim #3), we will test whether altered expression of known regulatory genes is involved in the atypical estrogen responsiveness of adult uteri in neonatally DES treated hamsters. We will begin with the c-myc c-tos and c-jun proto-oncogenes plus the p53 anti- oncogene. Their expression will be probed at the RVA and protein level as well as at the whole-organ and cell specific level using Northern blot analysis, in situ hybridization, immunoprecipitation/Western blot analysis and immunohistochemistry. Together these studies should contribute significantly to our long-term objectives of 1) understanding the basic mechanisms whereby estrogen regulates uterine growth and morphogenesis and 2) identifying mechanistic alterations that are responsible for degeneration of this process to the unregulated neoplastic state. These are biomedically important because 1) successful conception and gestation demands normal uterine form and function, and 2) estrogen-dependent uterine neoplasms ar responsible for considerable morbidity and mortality in contemporary American society.

当仓鼠在新生儿期被合成雌激素治愈时， 去乙雌酚 (UES)，其子宫始终表现出严重的 成年期对雌激素的增生/肿瘤反应。 两个之一 替代工作假设应该解释这种现象：1）直接 作用 新生仓鼠的细胞生理学和/或组成 子宫被 DES 损伤直接且永久地改变，使得 成人机体对雌激素的整体增殖反应变得不典型， 或 2) 间接作用 子宫营养活性由雌激素介​​导 主要通过或与其他不明因素结合，以及 新生儿 DES 治疗永久改变其水平或功能活动 等因素。 测试这些假设将开始（具体目标#1）： 监测对照和 DES 愈合的新生儿子宫的形态发生 不要将移植到对侧颊囊的动物 对照和 DES 处理的成熟宿主，已切除卵巢并使用雌激素 更换。 为了确认和扩展研究结果（具体目标#2），同型 以及对照子宫间质和上皮的异型重组 将进行 DES 处理的动物，并对其形态发生进行研究 研究：a) 在体外使用补充有子宫组织提取物的培养基 和/或来自特定目标#1和b)中使用的相同宿主组的血清 vivo（颊囊内）使用与特定目标中相同的主机组 #1. 因为这种方法的组合很少依赖于先前的经验 假设并适应体内和体外观察，坚定 应就哪种替代假设最有效得出结论 有效的。 最后（具体目标#3），我们将测试表达是否改变 已知的调节基因涉及非典型雌激素 新生 DES 治疗仓鼠成年子宫的反应性。 我们将 从 c-myc、c-tos 和 c-jun 原癌基因加上 p53 抗基因开始 癌基因。 它们的表达将在 RVA 和蛋白质水平上进行探测，如下所示 以及使用 Northern blot 在整个器官和细胞特异性水平 分析、原位杂交、免疫沉淀/蛋白质印迹分析 和免疫组织化学。 这些研究共同应该做出贡献 对我们的长期目标具有重要意义：1）了解基本知识 雌激素调节子宫生长和形态发生的机制 2）确定导致的机械改变 该过程退化为不受控制的肿瘤状态。 这些 具有生物医学重要性，因为 1) 成功受孕和妊娠 需要正常的子宫形态和功能，2) 雌激素依赖性 子宫肿瘤造成相当大的发病率和死亡率 在当代美国社会。