STERIOD REGULATED EXPRESSION IN THE UTERUS

大多数人类乳腺肿瘤中含有大量的 雌激素受体，以及大约三分之一的子宫内膜癌， 对某种形式的内分泌治疗有反应。然而，通过这种机制 激素在正常细胞或肿瘤细胞中控制细胞生长尚不清楚。 因此，我们建议从根本上调查这个问题。 女性生殖轨迹中的基因激活水平。我们的目标是 是为了创建一个mRNAs文库(一个cDNA库)，它是由 雌二醇在大鼠子宫内膜细胞中的表达。除 随机响应的mRNA将被克隆的鸟枪式方法，我们将 尝试丰富针对特定编码的消息的RNA制备 对于我们发现由子宫内膜细胞分泌的64K蛋白 对雌二醇的反应。然后，这些DNA将被用来探索动力学 诱导和去诱导对雌激素有反应的各种mRNAs 以及抗雌激素治疗和黄体酮的应用。我们会 检查它们在正常发情周期和发情早期的波动。 子宫发育。此外，我们将确定其中是否有任何 CDNAs探针与细胞系中发现的mRNAs发生交叉反应 来自人类子宫内膜癌(HEC-59)。最后，我们将尝试 确定这些mRNAs所来自的染色体DNA序列 抄写的。我们将用我们的cDNA依次探测大鼠基因组文库 寻找含有同源基因组插入片段的杂交噬菌体。这些插入物 将被映射和部分排序，目标是识别 可能与荷尔蒙有关的调控区域 在这组基因中可以看到控制力。

The majority of human mammary tumors that contain significant amounts of estrogen receptor, and about one-third of all endometrial carcinomas, respond to some form of endocrine therapy. Yet the mechanism by which hormones control cell growth in either normal or tumor cells is not known. We therefore propose an investigation into this problem at the fundamental level of gene activation in the female reproductive track. Our goal will be to create a library of mRNAs (a cDNA library) that are induced by estradiol in the endometrial cells of rat uterus. In addition to the shotgun approach in which random responsive mRNAs will be cloned, we will attempt to enrich a preparation of RNA for messages that code specifically for the 64K protein that we find to be secreted by endometrial cells in response to estradiol. These cDNAs will then be used to probe the kinetics of induction and deinduction of the various mRNAs that respond to estradiol and antiestrogen treatment and to progesterone administration. We will examine their fluctuation during the normal estrus cycle and during early uterine development. In addition, we will determine whether any of these cDNA probes cross-react with mRNAs that are found in a cell line derived from a human endometrial carcinoma (HEC-59). Finally, we will attempt to identify the chromosomal DNA sequences from which these mRNAs are transcribed. A rat genomic library will be probed with our cDNAs in order to find hybrid phages containing homologous genomic inserts. These inserts will be mapped and partially sequenced with the goal of identifying possible regulatory regions that may be responsible for the hormonal control seen in this set of genes.