NEONATAL DES-INDUCED UTERINE DYSPLASIA/NEOPLASIA

新生儿去诱导子宫发育不良/肿瘤

• 批准号: 3203946
• 负责人: WILLIAM J HENDRY
• 金额: $ 14.59万
• 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/3203946
• 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.

当用合成雌激素在新生儿时期治愈仓鼠时， 己烯雌酚(UES)，他们的子宫一致性表现出严重的 成年期雌激素的增生性/肿瘤性反应。两个中的一个 替代工作假说应该能解释这种现象：1)直接 对新生仓鼠细胞生理和/或成分的影响 子宫被DES侮辱直接和永久地改变 成年生物体对雌激素的整体增殖反应变得不典型， 或2)间接作用子宫的营养活性是由雌激素介导的 主要是通过或与其他不明因素相结合，以及 新生儿DES治疗永久性改变水平或功能活动 这样的因素。检验这些假设将从以下方面开始(具体目标#1) 对照和DES愈合后新生儿子宫形态发生的监测 不要将动物移植到对侧的脸颊囊中 对照和DES处理的成熟宿主去卵巢和雌激素- 被替换了。确认和扩展发现(具体目标2)，同型 对照组子宫间质和上皮细胞异型重组 将进行DES处理的动物，它们的形态发生将 研究：a)在体外使用添加子宫组织提取物的培养液 和/或来自在特定目标#1和b)中使用的相同主机组的血清 活体(在面颊袋内)使用与特定目标相同的主体组 #1.因为这种方法的组合依赖于很少的(如果有的话)先前 假设和适应体内和体外观察，坚定 应该得出关于哪种替代假设最多的结论 有效。最后(具体目标3)，我们将测试是否改变了表情 已知的调控基因与非典型雌激素有关 新生DES处理的仓鼠成年子宫的反应性。我们会 从c-myc、c-tos和c-jun原癌基因加上p53抗癌基因开始 致癌基因。它们的表达将在RVA和蛋白质水平上被检测为 以及在整个器官和细胞特异性水平上的Northern印迹 分析、原位杂交、免疫沉淀/Western印迹分析 和免疫组织化学。总之，这些研究应该有助于 对于我们的长期目标：1)了解基本的 雌激素调节子宫生长和形态发生的机制 2)确定导致以下问题的机械变更 这一过程退化为不受调控的肿瘤状态。这些 在生物医学上很重要，因为1)成功的受孕和怀孕 要求正常的子宫形态和功能，以及2)雌激素依赖 子宫肿瘤是相当大的发病率和死亡率的原因。 在当代美国社会。