NUCLEAR STEROID HORMONE RECEPTORS IN BREAST CANCER

由于两个原因，对孕激素的临床兴趣越来越大：一， 使用孕酮受体（PR）测量来标记 乳腺癌、子宫内膜癌、卵巢癌和前列腺癌; 第二，使用孕激素和抗孕激素避孕， 癌症的内分泌疗法。 我们已经开发出了一种化学计量的 PR的交换试验，并用它来显示孕酮可以 调节自身受体的水平， 雌激素 我们还表明，一个广泛使用的合成实验， 孕激素R5020对PR水平具有慢性抑制作用。 这个如果 外推到临床环境，表明PR患者服用 合成孕激素可能被错误地分配。 这些研究 这可能是因为永久性的人类乳腺癌细胞 株系，特别是一种称为T47 D的株系， 而不需要雌激素诱导。 我们在本申请中的目的是 详细研究，生物学，代谢和受体机制， 孕酮、合成孕激素和一种新的避孕抗孕激素 第486章. 我们计划使用T47 D细胞（雌激素非依赖性PR）MCF-7细胞 （雌激素依赖性PR）和BT-20细胞（PR阴性）：1）开发三种 生物学反应的标记resistin行动。 这些是抑制 细胞生长，胰岛素受体的诱导，以及分泌的 proteins. 2)我们将使用气相色谱和质谱来研究 并确定最终受体结合 负责生物活性的化合物。 3)我们计划研究 孕激素受体的分子生物学及其与孕激素的相互作用 通过DNA甲基化、组蛋白乙酰化和激素调节PR水平 抗性;全受体和核受体结构（色谱法） 和电泳; PR激活，易位核基质PR结合 免疫荧光位点;内切和外切核酸酶活性。 4)我们提出 用标记和未标记的方法研究抗孕酮作用的机制， 第486章.

There is increasing clinical interest in progestins for two reasons: one, the use of progesterone receptor (PR) measurements to mark hormone-dependent breast, endometrial, ovarian and prostatic cancers; and two, the use of progestins and antiprogestins for contraception and for endocrine therapies of cancer. We have developed a stoichiometric nuclear exchange assay for PR and have used it to show have progesterone can regulate the levels of its own receptors freed of the interfering effects of estrogens. We have also shown that a widely used synthetic experimental progestin, R5020, has chronic suppressive effects on PR levels. This, if extrapolated to the clinical setting, suggests that PR in patients taking synthetic progestins may be incorrectly assigned. These studies have been possible because of the availability of permanent human breast cancer cell lines, particularly one called T47D, that synthesizes enormous levels of PR without requiring estrogen induction. Our aim in this application is to study in detail, the biology, metabolism and receptor mechanisms of progesterone, synthetic progestins, and a new contraceptive antiprogestin RU38 486. We plan to use T47D cells (estrogen independent PR) MCF-7 cells (estrogen-dependent PR) and BT-20 cells (PR negative): 1) To develop three biological responses to mark progestin action. These are inhibition of cell growth, induction of insulin receptors, and synthesis of secreted proteins. 2) We will use gas chromatography and mass spectrometry to study cell mediated progestin metabolism and identify the ultimate receptor-bound compound responsible for biological activity. 3) We plan to study the molecular biology of PR and their interaction with progestins: the regulation of PR levels by DNA methylation, histone acetylation and hormone resistance; holoreceptor and nuclear receptor structures by chromatography and electrophoresis; PR activation, translocation nuclear matrix PR binding sites by immunofluorescence; endo and exonuclease activity. 4) We propose to study the mechanisms of antiprogestin action using labeled and unlabeled RU38 486.