Functional estrogen receptors in osteoblastic cells demonstrated by transfection with a reporter gene containing an estrogen response element.
Functional estrogen receptors in osteoblastic cells demonstrated by transfection with a reporter gene containing an estrogen response element.
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通过转染含有雌激素反应元件的报告基因来证明成骨细胞中的功能性雌激素受体。
作者：
Matthias Ernst;M. G. Parker;G. Rodan
虽然在成骨细胞中可见少量的雌激素受体（ER），并且雌二醇（E2）在体外对成骨细胞具有一定的作用，但E2对成骨细胞的直接作用尚未完全确定。为了确定成骨细胞中是否存在功能性ER，我们用含有氯霉素乙酰转移酶（CAT）报告基因和来自卵黄蛋白原A2基因的雌激素反应元件（ERE）的质粒转染细胞。在瞬时转染和随后用10-100 nM 17 β-E2处理后48 h测定CAT活性的E2依赖性诱导。17 β-E2，但不是17 α-E2，二氢睾酮，或孕酮，诱导CAT活性的剂量依赖性的方式（高达6倍）在大鼠颅骨部分-3，RCT-3，PyMS，和UMR-106细胞以及在人骨肉瘤细胞系SaOS-2/B-10。相反，E2对前成骨细胞系RCT-1和TRAB-11、大鼠骨肉瘤细胞系ROS 17/2.8和成纤维细胞系BALB-c/3 T3和NRK中CAT活性的诱导没有影响。使用猴病毒-40为基础的表达载体的ER的过度表达不仅赋予或增强E2依赖性诱导CAT在所有细胞类型，但增强E2依赖性表达的胰岛素样生长因子-I和E2刺激的DNA合成在原代颅骨和PyMS成骨细胞，分别。这些数据表明，存在低水平的功能性内源性雌激素受体在一些，但不是所有的，成骨细胞，并表明，丰富的雌激素受体可能是速率限制在E2对这些细胞的作用。
Although a small number of estrogen receptors (ER) were visualized in osteoblastic cells, and estradiol (E2) has some effects on osteoblasts in vitro, the direct action of E2 on osteoblasts has not been fully established. To determine the presence of functional ER in osteoblasts, we transfected cells with a plasmid containing the chloramphenicol acetyl transferase (CAT) reporter gene and the estrogen-responsive element (ERE) from the vitellogenin A2 gene. E2-dependent induction of CAT activity was determined 48 h after transient transfection and subsequent treatment with 10-100 nM 17 beta-E2. 17 beta-E2, but not 17 alpha-E2, dihydrotestosterone, or progesterone, induced CAT activity in a dose-dependent manner (up to 6-fold) in rat calvarial fraction-3, RCT-3, PyMS, and UMR-106 cells as well as in the human osteosarcoma cell line SaOS-2/B-10. In contrast, E2 had no effect on the induction of CAT activity in the preosteoblastic cell lines RCT-1 and TRAB-11, in the rat osteosarcoma cell line ROS 17/2.8, and in the fibroblastic cell lines BALB-c/3T3 and NRK. Over-expression of ER using a simian virus-40-based expression vector not only conferred or enhanced E2-dependent induction of CAT in all cell types, but augmented E2-dependent expression of insulin-like growth factor-I and E2-stimulated DNA synthesis in primary calvarial and PyMS osteoblastic cells, respectively. These data show the presence of low levels of functional endogenous ER in some, but not all, osteoblastic cells and suggest that the abundance of ER may be rate limiting in the action of E2 on these cells.
雌激素和抗雌激素的非基因组效应。
DOI：
10.1016/0022-4731(88)90017-9
发表时间：
1988
期刊：
Journal of steroid biochemistry
影响因子：
--
作者：
Weiss,DJ;Gurpide,E
通讯作者：
Gurpide,E
雌二醇直接调节人骨细胞 (SaOS-2) 对人甲状旁腺激素 (PTH) 和 PTH 相关蛋白的反应。
影响因子：
4.8
作者：
Fukayama,S;TashjianJr,AH
通讯作者：
TashjianJr,AH