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利用¹²⁵I-雌二醇放射自显影术对细胞和组织切片中的雌激素受体进行快速灵敏检测。

Rapid and sensitive detection of oestrogen receptors in cells and tissue sections by autoradiography with 125I-oestradiol.

作者信息

Berns E M, Rommerts F F, Mulder E

出版信息

Histochem J. 1985 Nov;17(11):1185-96. doi: 10.1007/BF01002501.

Abstract

The presence of receptors for steroid hormones in individual cells and tissue sections was assessed within 4-24 h using dry mount autoradiography with radio-iodinated oestradiol. Low affinity and nonspecific binding of steroids were significantly reduced by washing the cells or sections with diluted antiserum to oestradiol. For cells of the MCF-7 cell line variations in grain density were observed, indicating that cells of the MCF-7 cell line are heterogeneous with respect to their cellular receptor concentrations of oestrogen receptors. Receptor-negative cells, such as peritoneal macrophages, did not retain oestradiol label. In tissue sections of rat and calf uterus, predominant labelling was observed on the endometrial gland cells and stroma. Oestradiol receptor binding in the uterus cytosol for both radio-iodinated and tritiated oestradiol showed the same qualitative characteristics as determined by sucrose gradient sedimentation profiles and a comparable amount of binding sites was found for both labels. The relative binding affinity of 125I-oestradiol compared to [3H]oestradiol is about 70-80%. The dry mount autoradiographic technique as presented can be used for rapid screening of heterogeneity in oestrogen receptor distribution in cells and tissue sections, since this technique reveals differences in receptor concentrations on the single cell level.

摘要

在4至24小时内,使用放射性碘化雌二醇的干板放射自显影技术评估单个细胞和组织切片中类固醇激素受体的存在情况。用稀释的抗雌二醇血清洗涤细胞或切片可显著降低类固醇的低亲和力和非特异性结合。对于MCF - 7细胞系的细胞,观察到颗粒密度存在差异,这表明MCF - 7细胞系的细胞在雌激素受体的细胞受体浓度方面存在异质性。受体阴性细胞,如腹膜巨噬细胞,不保留雌二醇标记。在大鼠和小牛子宫的组织切片中,主要在子宫内膜腺细胞和基质上观察到标记。放射性碘化和氚化雌二醇在子宫胞浆中的雌二醇受体结合显示出与蔗糖梯度沉降谱测定相同的定性特征,并且两种标记物的结合位点数量相当。与[3H]雌二醇相比,125I - 雌二醇的相对结合亲和力约为70 - 80%。所介绍的干板放射自显影技术可用于快速筛选细胞和组织切片中雌激素受体分布的异质性,因为该技术可在单细胞水平揭示受体浓度的差异。

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