Ikegami A, Inoue S, Hosoi T, Mizuno Y, Nakamura T, Ouchi Y, Orimo H
Department of Geriatrics, Faculty of Medicine, University of Tokyo, Japan.
J Bone Miner Res. 1993 Sep;8(9):1103-9. doi: 10.1002/jbmr.5650080911.
Expression of estrogen receptor (ER) was studied in MC3T3-E1 cells (mouse osteoblastic cell line), HOS TE85 cells (human osteosarcoma cell line), and primary osteoblastic cells derived from mouse calvaria with immunohistochemical techniques. The staining of ER was readily detectable in MC3T3-E1 cells, HOS TE85 cells, and primary osteoblastic cells by using a monoclonal anti-ER antibody that recognizes the DNA binding domain of ER. The immunoreactivity was distributed in the cytoplasm as well as in the nuclei. 17 beta-Estradiol (10(-8) M) did not alter this staining pattern. The expression of ER was confirmed by Northern blot analysis using rat ER cDNA probe, which revealed a 6.5 kb band in MC3T3-E1 cells and a 6.2 kb band in HOS TE85 cells. The mRNA level of ER was not altered by 17 beta-estradiol (10(-8) M). The immunohistochemical studies showed that ER was not detectable in all cells but in a small population of each cell type. This study is the first report to demonstrate the presence of ER immunohistochemically, and our results suggest the heterogeneity of ER expression among osteoblastic cells.
采用免疫组织化学技术，在MC3T3-E1细胞（小鼠成骨细胞系）、HOS TE85细胞（人骨肉瘤细胞系）以及源自小鼠颅骨的原代成骨细胞中研究雌激素受体（ER）的表达。通过使用识别ER DNA结合域的单克隆抗ER抗体，在MC3T3-E1细胞、HOS TE85细胞和原代成骨细胞中很容易检测到ER的染色。免疫反应性分布于细胞质和细胞核中。17β-雌二醇（10⁻⁸ M）并未改变这种染色模式。使用大鼠ER cDNA探针通过Northern印迹分析证实了ER的表达，该分析显示在MC3T3-E1细胞中有一条6.5 kb的条带，在HOS TE85细胞中有一条6.2 kb的条带。17β-雌二醇（10⁻⁸ M）未改变ER的mRNA水平。免疫组织化学研究表明，并非所有细胞中都能检测到ER，而是在每种细胞类型的一小部分细胞中可检测到。本研究是首次通过免疫组织化学证明ER存在的报告，我们的结果提示成骨细胞中ER表达具有异质性。
