THE OVARY - A TARGET ORGAN FOR 1,25-DIHYDROXYVITAMIN-D3
THE OVARY - A TARGET ORGAN FOR 1,25-DIHYDROXYVITAMIN-D3
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DOI：
10.1210/endo-112-1-200
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发表时间：
1983-01-01
期刊：
影响因子：
4.8
通讯作者：
HAUSSLER, MR
中科院分区：
文献类型：
--
作者：
DOKOH, S;DONALDSON, CA;HAUSSLER, MR
使用培养的克隆中国仓鼠卵巢 (CHO) 细胞和母鸡卵巢，研究了特定 1,25-二羟基维生素 D3 [1,25-(OH)2D3] 受体的存在。对最初在低盐缓冲液中分离的 CHO 细胞的高盐胞质和母鸡卵巢核的高盐提取物进行了受体分析。 CHO 和母鸡卵巢均含有特定的高亲和力 1,25-(OH)2D3 受体 (Kd = 10-10-10-11 M)，其在高盐蔗糖梯度中的沉积 (3.3S) 与小鸡肠道受体的 1,25-(OH)2D3 相同。这两个来源的 3.3S 大分子在 0.1 M KCl 下吸附到 DNA 纤维素上，并在 0.2-0.22 M KCl 的线性盐梯度下洗脱，这是 1,25-(OH)2D3 受体的特性特征。饱和度分析表明有.apprx。每个 CHO 细胞有 2000 个受体分子拷贝。研究了 1,25-(OH)2D3 对 CHO 细胞单层培养物中卵巢细胞生长的影响。当培养基中存在生理 (100 pM) 水平的 1,25-(OH)2D3 时，观察到对 CHO 细胞生长的显着抑制（高达 60%）。这种生长抑制是剂量依赖性的，并且伴随着总细胞蛋白的平行减少。浓度高达 1 nM 的 25-羟基维生素 D3 和 24,25-二羟基维生素 D3 不会影响 CHO 细胞生长，这表明，与受体结合一样，1,25-(OH)2D3 比其他维生素 D 代谢物选择性地影响细胞增殖。哺乳动物和鸟类的卵巢细胞具有 1,25-(OH)2D3 受体，该受体可能在 1,25-(OH)2D3 对培养中这些细胞生长的影响中发挥作用。
Using both clonal Chinese hamster ovary (CHO) cells in culture and the hen ovary, a search was made for the presence of specific 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] receptors. Receptor analyses were carried out on high salt cytosols of CHO cells and high salt extracts of hen ovarian nuclei that were originally isolated in low salt buffer. Both CHO and hen ovary contain a specific high affinity 1,25-(OH)2D3 receptor (Kd = 10-10-10-11 M), which sediments (3.3S) in high salt sucrose gradients identically to the chick intestinal receptor for 1,25-(OH)2D3. This 3.3S macromolecule from both sources adsorbed to DNA-cellulose at 0.1 M KCl and eluted during a linear salt gradient at 0.2-0.22 M KCl, a property characteristic of the 1,25-(OH)2D3 receptor. Saturation analysis indicated that there are .apprx. 2000 copies of the receptor molecule per CHO cell. The effect of 1,25-(OH)2D3 on ovarian cell growth in monolayer culture of CHO cells was investigated. Significant inhibition of CHO cell growth (up to 60%) was observed in the presence of physiological (100 pM) levels of 1,25-(OH)2D3 in the culture medium. This inhibition of growth was dose dependent and was accompanied by a parallel decrease in total cell protein. Concentrations of 25-hydroxyvitamin D3 and 24,25-dihydroxyvitamin D3 as high as 1 nM did not affect CHO cell growth, indicating that, like receptor binding, cell proliferation is selectively influenced by 1,25-(OH)2D3 over other vitamin D metabolites. Ovarian cells in mammals and birds possess the 1,25-(OH)2D3 receptor which may play a role in the effect of 1,25-(OH)2D3 on the growth of these cells in culture.