MECHANISM OF ESTROGEN ACTION IN UTERUS AND VAGINA

雌激素17-β(E_2)和孕酮(P)是主要的激素 上皮细胞增殖、形态发生的调节， 阴道和子宫的细胞分化和分泌活动。 E2和P在这些器官中的作用是通过雌激素介导的 受体(ER)和孕激素受体(PR)。呃和 PR位于子宫上皮细胞和间质细胞中， 阴道，但有证据表明，这些药物的某些作用 子宫和阴道上皮激素可能是间接介导的 通过间质ER和PR。在本提案中，我们将利用 雌激素受体基因敲除(ERKO)小鼠联合组织 分离/重组技术生产子宫和阴道 在基质和上皮中都表达ER的组织重组体，在 仅间质或上皮，或在这两种类型的细胞中都不存在。 这些组织重组体随后将在体内移植，并用于 直接测定雌二醇对子宫上皮细胞的影响 分泌蛋白产生、细胞凋亡和孕激素受体上调- 调节是通过上皮或间质中的内质网来实现的。这个 表皮生长因子受体信号在间质和上皮组织中的作用 还将确定E2的调节作用。阴道组织 用对照和ERKO组织制备的重组体将用于 确定上皮内质网在阴道层积中的作用， 角化和粘液化。类似的实验将会进行。 利用孕激素受体基因敲除(PRKO)小鼠的组织 确定P对阴道粘液和雌二醇诱导的作用 子宫上皮细胞的增殖是通过间质或 上皮性PR。这项工作对于理解 雌二醇和孕酮正常调控发育的基本机制 阴道和子宫上皮中发生的事件。此外，这项工作是 对子宫内膜异位症等疾病具有潜在的重要临床意义， 它涉及子宫上皮的异常增殖，对于 女性生殖道癌，几乎完全是上皮性癌 起源，因为E2似乎至少在这些 疾病和P可能拮抗E_2的作用。

Estradiol 17-beta (E2) and progesterone (P) are the major hormonal regulators of epithelial proliferation, morphogenesis, cytodifferentiation and secretory activity in the vagina and uterus. E2 and P effects in these organs are mediated through estrogen receptors (ER) and progesterone receptors (PR), respectively. ER and PR are located in both epithelial and stromal cells of the uterus and vagina, but some evidence indicates that certain effects of these hormones on uterine and vaginal epithelium may be mediated indirectly through stromal ER and PR. In this proposal, we will utilize the estrogen receptor knockout (ERKO) mouse in conjunction with tissue separation/recombination techniques to produce uterine and vaginal tissue recombinants that express ER in both stroma and epithelium, in only the stroma or the epithelium, or in neither of these cells types. These tissue recombinants will then be grafted in vivo, and used to directly determine whether the effects of E2 on uterine epithelial secretory protein production, apoptosis and progesterone receptor up- regulation are mediated through ER in the epithelium or stroma. The role of EGF receptor signalling in stromal and epithelial tissue for mediating the effects of E2 will also be determined. Vaginal tissue recombinants prepared with control and ERKO tissue will be used to determine the role of epithelial ER in vaginal stratification, cornification and mucification. Similar experiments will be performed using tissue from progesterone receptor knockout (PRKO) mice to determine if the effects of P on vaginal mucification and E2-induced uterine epithelial proliferation are mediated through stromal or epithelial PR. This work will be of vital importance for understanding the basic mechanisms by which E2 and P normally control developmental events in vaginal and uterine epithelium. Additionally, this work is potentially important clinically for diseases such as endometriosis, which involves aberrant proliferation of uterine epithelium, and for female reproductive cancers, which are almost exclusively of epithelial origin, since E2 appears to act at least as a permissive agent in these diseases and P may antagonize the effects of E2.