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

雌激素靶细胞中的原癌基因和生长因子/受体活性

• 批准号: 6239905
• 负责人: WINSTON A ANDERSON
• 金额: $ 4.83万
• 依托单位: HOWARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-01 至 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6239905
• 关键词: athymic mouse; autoradiography; cell cycle; cell differentiation; estrogen receptors; estrogens; genetic transcription; growth factor; growth factor receptors; high performance liquid chromatography; hormone receptor; in situ hybridization; laboratory rat; messenger RNA; northern blottings; nuclear runoff assay; peroxidases; protein biosynthesis; protooncogene; receptor binding; tamoxifen; uterus; western blottings

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.

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的肿瘤细胞和组织中。 雌激素对多种核基因的诱导、抑制和抑制 已知参与这些增殖性和分化的事件。多么 抗雌激素，如他莫昔芬，调节雌激素的早期事件 归纳法完全不确定。因此，主要目标是 这项研究将探讨雌激素的调节作用和 三苯氧胺对已知在增殖中起关键作用的特定基因的影响 和/或啮齿动物子宫组织中的分化。具体地说， 这些研究中使用的探针用于评估增殖和 分化将包括生长因子(EGF、IGF-1、IGF-2外显子2、 IGF-2外显子4，转化生长因子β1，2，3型)，生长因子受体(EGFR，IGF- 1R、IGF-2R、TGFbeta-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 在0‘，15，30’，1小时， 6小时、24小时、48小时和72小时。表情研究将由以下人员分析 斑点/Northern杂交，以及与上述蛋白的斑点杂交 探测器。放射自显影将通过密度扫描进行分析。在……里面 相关研究我们将确定过氧化物酶基因是否在 直接控制由E2-ER复合体激活的基因产物。至 回答问题，点/槽研究使用来自 基因组文库将用于鉴定雌激素诱导的过氧化物酶 (EIP)mRNA。克隆的与EIP mRNA同源的DNA片段将 测序并与子宫染色质杂交，试图 对EIP基因进行定位。