IMMUNOELECTRONMICROSCOPIC LOCALIZATION OF A PROLIFERATION-ASSOCIATED ANTIGEN KI-67 IN MCF-7 CELLS
IMMUNOELECTRONMICROSCOPIC LOCALIZATION OF A PROLIFERATION-ASSOCIATED ANTIGEN KI-67 IN MCF-7 CELLS
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DOI：
10.1007/bf01007235
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发表时间：
1990-09-01
期刊：
影响因子：
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通讯作者：
KALLIONIEMI, OP
中科院分区：
文献类型：
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作者：
ISOLA, J;HELIN, H;KALLIONIEMI, OP
使用单克隆抗体Ki-67的免疫细胞化学是评估恶性肿瘤增殖活性的常用方法。Ki-67与具有抗原的增殖细胞反应，其结构、功能和确切位置尚不清楚。我们研究了Ki-67在MCF-7细胞中的亚细胞定位，使用免疫电镜。在间期细胞中，Ki-67免疫反应位于核仁，主要位于核仁皮质。特别是，核仁的颗粒成分的区域被强烈染色。在核仁外也可见弱点状核质免疫染色。在前期Ki-67抗原定位在凝聚的染色质的表面上，在中期在染色体的表面上。细胞分裂后，新的核仁形成之前，Ki-67免疫反应位于核质。通过流式细胞术定量Ki-67免疫荧光信号显示有丝分裂细胞中Ki-67水平最高。Ki-67的位置是非常相似的某些最近描述的蛋白质的核仁preribosomes表明Ki-67也可能是一个组成部分的preribosomes。
Immunocytochemistry using the monoclonal antibody Ki-67 is a commonly used method to assess proliferative activity of malignant tumours. Ki-67 reacts with proliferating cells with an antigen, whose structure, function and exact locations are unknown. We studied the subcellular location of Ki-67 in MCF-7 cells using immunoelectron microscopy. In the interphase cells, Ki-67 immunoreactivity was localized in the nucleolus, mainly in the nucleolar cortex. In particular, areas of the granular component of the nucleolus were strongly stained. Weak spot-like nucleoplasmic immunostaining was also seen outside the nucleoluse. During prophase Ki-67 antigen was localized on the surfaces of the condensed chromatin and during metaphase on the surface of the chromosomes. After cell division and prior to formation of new nucleoli, Ki-67 immunoreactivity was located in the nucleoplasm. Quantification of Ki-67 immunofluorescence signal by flow cytometry revealed highest Ki-67 levels in mitotic cells. The location of Ki-67 is very similar to certain recently described proteins of nucleolar preribosomes suggesting that Ki-67 may also be a component of the preribosomes.