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首页> 外文期刊>International Journal of Clinical and Experimental Pathology >Differential effects of estrogen and estrogen receptor antagonist, ICI 182 780, on the expression of calbindin-D9k in rat pituitary prolactinoma GH3 cells
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Differential effects of estrogen and estrogen receptor antagonist, ICI 182 780, on the expression of calbindin-D9k in rat pituitary prolactinoma GH3 cells

机译：雌激素和雌激素受体拮抗剂ICI 182780对大鼠垂体泌乳素瘤GH3细胞中calbindin-D9k表达的影响

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Objective: To detect the effects of 17β-estradiol (E2) on the expression of calbindin-D9k (CaBP-9k) in pituitary GHsub3/sub cells, and to determine the antagonistic effect of a selective estrogen receptor (ER) antagonist (ICI 182 780) on CaBP-9k expression. Methods: A rat pituitary prolactinoma cell line (GHsub3/sub cells) was used in an emin vitro/em model. The localization of CaBP-9k in GHsub3/sub cells was observed by immunofluorescence. GHsub3/sub cells were cultured with the addition of E2 medium for 24 hours. The levels of CaBP-9k mRNA and protein expression in different groups were analyzed by RT-PCR and Western blot analysis. The ER antagonist, ICI 182 780, was added to GHsub3/sub cells before E2 (10sup-8/sup M) at a concentration of 10sup-6/sup M to investigate the regulation of an ER-mediated pathway on CaBP-9k expression. Results: E2 had a stimulatory effect on CaBP-9k expression of GHsub3/sub cells in a dose-dependent manner; the level of CaBP-9k expression was higher when treated with a higher concentration of E2. ICI 182 780 suppressed the stimulatory effect of E2 on CaBP-9k expression in GHsub3/sub cells. The level of CaBP-9k expression was significantly reduced by co-administration of E2 with ICI 182 780 in GHsub3/sub cells. The immunoprecipitation results confirmed that CaBP-9k interacts directly with ERα, and E2 increases the interaction between CaBP-9k and ERα. Conclusion: Estrogen induces CaBP-9k expression via an ERα-mediated pathway and CaBP-9k directly combines with ERα, suggesting that CaBP-9k is involved in the biological effects mediated by an ER pathway in GHsub3/sub cells.
机译：目的：检测17-雌二醇（E2）对垂体GH 3 细胞钙结合蛋白-D9k（CaBP-9k）表达的影响，并确定其选择性拮抗作用。雌激素受体（ER）拮抗剂（ICI 182 780）对CaBP-9k的表达。方法：将大鼠垂体泌乳素瘤细胞系（GH 3 ）用于体外模型。免疫荧光观察CaBP-9k在GH 3 细胞中的定位。加入E2培养基培养GH 3 细胞24小时。通过RT-PCR和Western blot分析分析不同组中CaBP-9k mRNA和蛋白表达水平。在E2（10 -8 M）之前将ER拮抗剂ICI 182780加入到GH 3 细胞中，浓度为10 -6 M以研究ER介导的途径对CaBP-9k表达的调控。结果：E2对GH 3 细胞CaBP-9k的表达具有剂量依赖性。当用较高浓度的E2处理时，CaBP-9k的表达水平较高。 ICI 182780抑制了E2对GH 3 细胞中CaBP-9k表达的刺激作用。 E2与ICI 182780在GH 3 细胞中共同施用可显着降低CaBP-9k的表达水平。免疫沉淀结果证实，CaBP-9k与ER＆＃x003b1直接相互作用，而E2增加CaBP-9k与ER＆＃x003b1;之间的相互作用。结论：雌激素通过ER介导的途径诱导CaBP-9k表达，而CaBP-9k直接与ER介导结合，表明CaBP-9k参与了GH的ER途径介导的生物学效应。 3 个单元格。

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