PROTO-ONCOGENE AND GROWTH FACTOR/RECEPTOR ACTIVITIES IN ESTROGEN TARGET CELLS

复杂的激素、生长因子和互补受体相互作用 调节正常循环中的增殖和分化变化 子宫组织和雌激素依赖性肿瘤细胞和组织中。 雌激素对多种核基因的诱导、阻遏和抑制作用 已知介导这些增殖和分化事件。 如何 抗雌激素，如他莫昔芬，调节雌激素的早期事件， 归纳是完全不确定的。 因此， 本研究将探讨雌激素的调节作用， 他莫昔芬对已知在增殖中起关键作用的特定基因的作用 和/或分化。 具体而言是 在这些研究中使用的探针来评估增殖， 分化将包括生长因子（EGF，IGF-1，IGF-2外显子2， IGF-2外显子4，TGF β 1、2、3型），生长因子受体（EGFR，IGF- 1 R，IGF-2 R，TGF β-2R，3R），激素受体（ER，hPR和原癌基因 （c-myc; c-fos; c-ras; c-Jun）。 在这项时间研究中，polyA+ RNA将 从雌激素和他莫昔芬处理的子宫中在0 '，15，30'，1小时， 6小时、24小时、48小时和72小时。 表达研究将通过以下方法进行分析： 斑点/北方印迹和斑点印迹/杂交 probes. 将通过光密度扫描分析放射自显影图。 在 相关的研究，我们将确定是否过氧化物酶基因下， 直接控制E2-ER复合物激活的基因产物。 到 回答这个问题，点/槽研究使用重组DNA克隆从 基因组文库将用于鉴定雌激素诱导的过氧化物酶 (EIP)mRNA。与EIP mRNA同源的克隆DNA片段将被 测序并与子宫染色质杂交， 定位EIP基因。

Complex hormonal, growth factor and complimentary receptor interactions regulate proliferative and differentiative changes in normal cycling uterine tissues and in estrogen-dependent tumor cells and tissues. Estrogen induction, repression and depression of various nuclear genes are known to mediate these proliferative and differentiative events. How antiestrogens, like Tamoxifen, regulate early events in estrogen induction is totally undetermined. Consequently, the major objective of this study will be to investigate the regulatory effects of estrogen and Tamoxifen on specific genes known to play key roles in proliferation and/or differentiation in rodent uterine tissue. Specifically, the probes used in these studies to evaluate proliferation and differentiation will include growth factors (EGF, IGF-1, IGF-2 Exon 2, IGF-2 Exon 4, TGFBeta-types 1,2,3), growth factor receptors (EGFR, IGF- 1R, IGF-2R, TGFBeta-2R,3R), hormone receptors (ER,hPR and protooncogenes (c-myc; c-fos; c-ras; c-Jun). In this temporal study, polyA+ RNA will be isolated from estrogen and Tamoxifen-treated uteri at 0',15,30', 1hr, 6hr, 24hr, 48hr, and 72hr. Expression studies will be analyzed by dot/Northern blot, and dot blot/hybridization with the above mentioned probes. Autoradiograms will be analyzed by densitometric scans. In related studies we will determine if the peroxidase gene is under the direct control of gene products activated by the E2-ER complex. To answer this question, dot/slot studies using recombinant DNA clones from a genomic library will be used to identify estrogen-induced peroxidase (EIP) mRNAs. Cloned DNA fragments homologous with the EIP mRNA will be sequenced and hybridized to chromatin from uteri in an attempt to localize the EIP gene.