MECHANISM OF HORMONE ACTION ON TARGET CELLS IN CULTURE

这项提案的长期目标是机械地定义 正常组织和肿瘤组织中细胞增殖的调节。人血清 抑制对雌激素敏感的人乳腺癌细胞的增殖； 雌激素以一种特定的方式克服了这种影响。其他性激素 而生长因子在逆转这种抑制作用方面完全无效。 这种抑制作用似乎是由于一种热稳定的、对蛋白酶敏感的 表观分子质量为70-90 kDa，等电性 打印4.5-4.8。此应用程序的具体目标是进一步 使用各种程序提纯该特定的抑制物，包括： 疏水相互作用和螯合层析，制备性 等电聚焦、凝胶过滤、二维聚丙烯酰胺凝胶 电泳法、高效液相色谱联用法及单抗和多克隆抗体检测。这个 用生物测定法测定纯化组分的抑制性能 利用MCF7人乳腺肿瘤细胞。 这些生理学的血管紧张素转换酶抑制剂的潜在临床意义 乳腺癌细胞的增殖是多方面的。其中包括以下方法： 更准确地预测乳腺肿瘤对治疗的反应 读取组织病理学样本时所做的决定；受体 血浆携带的抑制剂有望成为更可靠的标记物 比现在使用的探针(雌激素受体等)更能预测预后。同样 重要的是，我们正在提纯的抑制物的可用性将允许 对现在提出的解释控制的假设进行检验 雌激素敏感的细胞增殖。

The long term objective of this proposal is to define mechanistically the regulation of cell proliferation in normal and tumor tissues. Human serum inhibits the proliferation of estrogen-sensitive human breast cancer cells; estrogens overcome this effect in a specific manner. Other sex steroids and growth factors are completely ineffective in reversing this inhibition. The inhibitory effect seems to be due to a heat-stable, protease-sensitive molecule of an apparent molecular weight = 70-90 kDa and an isoelectric print of 4.5-4.8. The specific aim of this application is to further purify this specific inhibitor using a variety of procedures including: hydrophobic interaction and chelating chromatography, preparative isoelectric focusing, gel filtration, 2 dimensional polyacrylamide gel electrophoresis, HPLC/FPLC and mono and polyclonal antibodies. The bioassay to measure the inhibitory properties of the purified fractions makes use of MCF7 human breast tumor cells. The potential clinical relevance of these physiologic inhibitors of the proliferation of breast cancer cells are manifold. They include ways to more accurately prognosticate the response of breast tumors to therapeutic decisions taken when histopathological samples are read; receptors for the plasma-borne inhibitors are anticipated to be a more reliable marker of prognosis than probes used now (estrogen receptor, etc.). Equally important, availability of the inhibitory we are purifying, will allow for testing of hypotheses now being proposed to explain the control of estrogen-sensitive cell proliferation.