Simpson E R, Ackerman G E, Smith M E, Mendelson C R
Proc Natl Acad Sci U S A. 1981 Sep;78(9):5690-4. doi: 10.1073/pnas.78.9.5690.
Stromal cells prepared from adipose tissue of women were maintained in monolayer culture to study the regulation of aromatase activity by hormones. Aromatase activity was stimulated 20- to 100-fold by dexamethasone at a concentration of 250 nM. Half-maximal stimulation of aromatase activity was attained at a dexamethasone concentration of 2.7 nM. The stimulatory effect of dexamethasone was apparent after a preincubation time of 4 hr, and stimulation was maximal after 24 hr of preincubation. The stimulatory effect of dexamethasone was observed only when fetal calf serum also was present in the culture medium. Of the various steroids tested, dexamethasone was the most potent in stimulating aromatase activity. Cortisol was less effective than dexamethasone, whereas corticosterone, at a concentration of 250 nM, caused only a small stimulation of aromatase activity. Progesterone and deoxycorticosterone (250 nM) did not affect aromatase activity. Cytosolic fractions prepared from stromal cells that had been maintained in monolayer culture were found to contain a homogenous population of sites that specifically bound [3H]dexamethasone with relatively high affinity (Kd = 2.9 nM) and low capacity (38 fmol per mg of protein). The stimulatory effect of dexamethasone on aromatase activity was prevented by simultaneous incubation with cortisol 21-mesylate (0.1-10 microM), a compound known to block the binding of glucocorticosteroids to cytoplasmic receptors. The stimulatory effect of dexamethasone also was prevented by incubation of the cells with cycloheximide or actinomycin D. These findings are suggestive that glucocorticosteroids act to increase aromatase activity in stromal cells by inducing the synthesis of new enzyme protein.
从女性脂肪组织制备的基质细胞进行单层培养,以研究激素对芳香化酶活性的调节作用。在浓度为250 nM时,地塞米松可使芳香化酶活性增强20至100倍。在2.7 nM的地塞米松浓度下可达到芳香化酶活性的半数最大刺激。在预孵育4小时后,地塞米松的刺激作用明显,预孵育24小时后刺激作用达到最大。仅当培养基中也存在胎牛血清时,才观察到地塞米松的刺激作用。在测试的各种类固醇中,地塞米松在刺激芳香化酶活性方面最有效。皮质醇的效果不如地塞米松,而在浓度为250 nM时,皮质酮仅引起芳香化酶活性的轻微刺激。孕酮和脱氧皮质酮(250 nM)不影响芳香化酶活性。发现从单层培养的基质细胞制备的胞质部分含有一群均一的位点,这些位点以相对高的亲和力(Kd = 2.9 nM)和低容量(每毫克蛋白质38 fmol)特异性结合[3H]地塞米松。同时与21-甲磺酸皮质醇(0.1 - 10 microM)一起孵育可阻止地塞米松对芳香化酶活性的刺激作用,21-甲磺酸皮质醇是一种已知可阻断糖皮质激素与细胞质受体结合的化合物。用环己酰亚胺或放线菌素D孵育细胞也可阻止地塞米松的刺激作用。这些发现提示糖皮质激素通过诱导新的酶蛋白合成来增加基质细胞中的芳香化酶活性。