PROGESTERONE-RECEPTOR REGULATION IN UTERINE CELLS - STIMULATION BY ESTROGEN, CYCLIC ADENOSINE-3',5'-MONOPHOSPHATE, AND INSULIN-LIKE GROWTH FACTOR-I AND SUPPRESSION BY ANTIESTROGENS AND PROTEIN-KINASE INHIBITORS
PROGESTERONE-RECEPTOR REGULATION IN UTERINE CELLS - STIMULATION BY ESTROGEN, CYCLIC ADENOSINE-3',5'-MONOPHOSPHATE, AND INSULIN-LIKE GROWTH FACTOR-I AND SUPPRESSION BY ANTIESTROGENS AND PROTEIN-KINASE INHIBITORS
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DOI：
10.1210/endo-128-4-2045
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发表时间：
1991-04-01
期刊：
影响因子：
4.8
通讯作者：
KATZENELLENBOGEN, BS
中科院分区：
文献类型：
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作者：
ARONICA, SM;KATZENELLENBOGEN, BS
用原代培养的子宫细胞研究孕激素受体（PR）的多因素调控和可能介导这种调控的信号转导途径。 通过霍乱毒素加异丁基甲基黄嘌呤处理或加入8-溴-cAMP引起的细胞内cAMP增加，导致PR细胞内含量增加6至7倍，如通过[H-3]R5020结合和使用抗PR抗体的Western免疫印迹所监测。 在这些从19日龄未成熟大鼠分离的子宫细胞的原代培养物中，8-溴-cAMP在8 h时引起PR显著增加，在24 h时增加最大。 PR刺激的时间过程和幅度与最大有效浓度的雌二醇（3 × 10（-9）M）或IGF-I（20 ng/ml）诱发的PR刺激相似。 剂量反应研究表明，10（-6）至10（-4）M浓度的8-溴-cAMP（8-Br-cAMP）引起最大反应。 相比之下，在宽浓度范围内的8-溴-cGMP不能提高细胞PR水平。 在这些培养条件下，用这些试剂中的任何一种处理都不会改变细胞增殖。 虽然雌激素、cAMP和胰岛素样生长因子I（IGF-I）可能通过不同的途径起作用以增加PR，但这些药物的最大有效浓度引起的效应不是累加的，这意味着涉及共同的组分。 雌二醇、cAMP或IGF-I引起的PR升高可被抗雌激素治疗显著抑制（ICI 164，384）或环核苷酸依赖性蛋白激酶抑制剂H8或蛋白激酶A抑制剂PKI，表明雌激素受体和磷酸化途径参与这三种药物的PR调节。作为子宫细胞中PR水平的重要调节剂，并表明多种因素，包括影响细胞内cAMP水平的因素，可能通过调节细胞内PR含量来影响对孕激素的反应性。
Primary uterine cell cultures were used to study multifactor regulation of progesterone receptor (PR) and the signal transduction pathways which may serve to mediate that regulation. Increases in intracellular cAMP, brought about by treatment with cholera toxin plus isobutyl methyl xanthine or by addition of 8-bromo-cAMP, result in 6- to 7-fold increases in the intracellular content of PR as monitored by [H-3]R5020 binding and by Western immunoblot using anti-PR antibodies. In these primary cultures of uterine cells isolated from 19-day-old immature rats, 8-bromo-cAMP evokes significant increases in PR by 8 h with maximal increases by 24 h. This time course and magnitude of PR stimulations are similar to those evoked by maximally effective concentrations of estradiol (3 x 10(-9) M) or IGF-I (20 ng/ml). Dose-response studies reveal that 10(-6) to 10(-4) M concentrations of 8-bromo-cAMP (8-Br-cAMP) elicit a maximal response. In contrast, 8-bromo-cGMP over a wide concentration range was unable to elevate cellular PR levels. Under these culture conditions, cell proliferation was not altered by treatment with any of these agents. Although estrogen, cAMP, and insulin-like growth factor I (IGF-I) may act via different pathways to increase PR, the effects evoked by maximally effective concentrations of these agents are not additive implying involvement of a common component. The increases in PR evoked by estradiol, cAMP, or IGF-I are markedly suppressed by treatment with antiestrogen (ICI 164,384) or the cyclic nucleotide-dependent protein kinase inhibitor H8 or the protein kinase A inhibitor PKI, indicating the involvement of the estrogen receptor and phosphorylation pathways in PR regulation by these three agents.The present studies identify cAMP, as well as estrogen and IGF-I, as important regulators of the level of PR in uterine cells and suggest that multiple factors, including those affecting intracellular cAMP levels, might influence reponsiveness to progestins via regulation of the intracellular PR content.