Colston K W, King R J, Hayward J, Fraser D I, Horton M A, Stevenson J C, Arnett T R
St. George's Hospital Medical School, London, UK.
J Bone Miner Res. 1989 Aug;4(4):625-31. doi: 10.1002/jbmr.5650040421.
In this immunocytochemical study we have probed a number of human bone cell types and bone preparations for the presence of the estrogen receptor (ER) with two distinct monoclonal antibodies. Using a well-validated antibody (H222) that recognizes human ER and standard peroxidase-antiperoxidase methodology, we were unable to demonstrate nuclear staining for ER in cultured primary or transformed human bone-derived cells or in fetal bone sections. Attempts to visualize ER in osteosarcoma cell lines (TE85C and HTB96) using a silver enhancement procedure were also unsuccessful. Additionally, we failed to detect immunocytochemical staining for the progesterone receptor (using monoclonal antibody mPR1) in control or estrogen-treated human bone cell cultures. Estrogen and progesterone receptor staining was readily detectable in MCF7 human breast cancer cells. In contrast, with a monoclonal antibody that recognizes a 29 kDa cytoplasmic component (p29) closely related to human ER, we observed specific staining in all the osteoblastlike cells studied. Cytoplasmic staining for this p29 antigen was most intense in primary cultures of human bone-derived cells. It is possible that the relatively abundant but as yet undefined p29 antigen may act as a sensitive marker for the presence of ER in cells at levels below the detection limit of the anti-ER monoclonal antibody. If so, our results are consistent with the presence of ER in osteoblastlike cells at very low concentrations.
在这项免疫细胞化学研究中,我们使用两种不同的单克隆抗体,对多种人类骨细胞类型和骨标本进行雌激素受体(ER)检测。使用一种经过充分验证、能识别人类ER的抗体(H222)以及标准的过氧化物酶-抗过氧化物酶方法,我们无法在培养的原代或转化的人类骨源细胞或胎儿骨切片中证明ER的核染色。使用银增强程序在骨肉瘤细胞系(TE85C和HTB96)中观察ER的尝试也未成功。此外,在对照或雌激素处理的人类骨细胞培养物中,我们未能检测到孕激素受体的免疫细胞化学染色(使用单克隆抗体mPR1)。在MCF7人乳腺癌细胞中可轻易检测到雌激素和孕激素受体染色。相比之下,使用一种能识别与人类ER密切相关的29 kDa细胞质成分(p29)的单克隆抗体,我们在所研究的所有成骨样细胞中观察到特异性染色。这种p29抗原的细胞质染色在人类骨源细胞的原代培养物中最为强烈。相对丰富但尚未明确的p29抗原可能作为细胞中ER存在的敏感标志物,其水平低于抗ER单克隆抗体的检测限。如果是这样,我们的结果与成骨样细胞中存在极低浓度的ER一致。