Harmon J M, Thompson E B
J Cell Physiol. 1982 Feb;110(2):155-60. doi: 10.1002/jcp.1041100208.
Treatment of CEM-C7 cells with glucocorticoids produces a 2.5-fold increase in the activity of the enzyme glutamine synthetase (GS). This increase is specific for steroids with glucocorticoid activity adn occurs over a range of steroid concentrations consistent with a receptor-mediated mechanism. Half-maximal and maximal inductions by dexamethasone (dex) occur at 2 X 10(-8) M and 2 X 10(-7) M dex, respectively, concentrations approximately equal to those necessary to produce half and full occupancy of glucocorticoid receptors. GS activity began to increase 1 hour after dex treatment and was complete by 12 hours. This is well before any of the growth inhibitory or cytolytic effects of dex on this cell line occur. This increase was dependent on the presence of glucocorticoid receptors and required both RNA and protein synthesis. Removal of dex following stimulation to maximal levels resulted in a decrease of GS activity to preinduced levels with a half-time of 5 hours. Glutamine deprivation of cells resulted in increased GS activity. However, even in the total absence of glutamine, dex treatment elicited a 2.0-2.5-fold increase in GS activity, ruling out inhibition of glutamine uptake as a mechanism for the dex-induced increase. Experiments with 5'-bromodeoxyuridine (BrdU) demonstrated that GS elevation was sensitive to BrdU substitution of DNA, while dex-induced growth inhibition was not. Therefore GS elevation and growth inhibition in this cell line appear to be independently expressed steroid responses.
用糖皮质激素处理CEM - C7细胞会使谷氨酰胺合成酶(GS)的活性增加2.5倍。这种增加对具有糖皮质激素活性的类固醇具有特异性,并且在一系列与受体介导机制一致的类固醇浓度范围内发生。地塞米松(dex)诱导GS活性达到半数最大值和最大值时的浓度分别为2×10⁻⁸ M和2×10⁻⁷ M dex,这些浓度大致等同于使糖皮质激素受体达到半数占据和完全占据所需的浓度。地塞米松处理后1小时,GS活性开始增加,12小时时达到最大值。这远早于地塞米松对该细胞系产生任何生长抑制或细胞溶解作用的时间。这种增加依赖于糖皮质激素受体的存在,并且需要RNA和蛋白质合成。在刺激使GS活性达到最大水平后去除地塞米松,会导致GS活性下降至诱导前水平,半衰期为5小时。细胞缺乏谷氨酰胺会导致GS活性增加。然而,即使在完全没有谷氨酰胺的情况下,地塞米松处理仍能使GS活性增加2.0 - 2.5倍,排除了抑制谷氨酰胺摄取作为地塞米松诱导增加的机制。用5'-溴脱氧尿苷(BrdU)进行的实验表明,GS活性升高对地塞米松诱导的DNA中BrdU替代敏感,而地塞米松诱导的生长抑制则不敏感。因此,该细胞系中GS活性升高和生长抑制似乎是独立表达的类固醇反应。