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CD44两种亚型在人子宫内膜中的表达。

Expression of two isoforms of CD44 in human endometrium.

作者信息

Behzad F, Seif M W, Campbell S, Aplin J D

机构信息

Department of Obstetrics and Gynaecology, University of Manchester, United Kingdom.

出版信息

Biol Reprod. 1994 Oct;51(4):739-47. doi: 10.1095/biolreprod51.4.739.

Abstract

The distribution of the cell-surface adhesion glycoprotein CD44 in human endometrium was examined by immunofluorescence using six monoclonal antibodies to epitopes common to all forms of the molecule, and by reverse transcription-polymerase chain reaction (RT-PCR). Immunoreactivity was observed throughout the menstrual cycle in stroma, vessels, glandular, and luminal epithelium. Variations in staining intensity were observed, especially in the epithelial compartment. CD44 was also expressed strongly by decidualized stromal cells of first-trimester pregnancy. No systematic variation of immunoreactivity was observed with stages of the normal cycle, but a fraction (25%) of the specimens lacked reactivity in the epithelium. To determine the molecular size of the epithelial isoform, an immunoprecipitation technique was developed using surface-radioiodinated, detergent-extracted glands. This indicated the presence at the cell surface of a single dominant CD44E species with an approximate molecular mass of 130 kDa. RT-PCR was used to investigate the isoforms present in whole endometrial tissue, isolated gland fragments, and Ishikawa endometrial carcinoma cells. Complementary DNA produced from total endometrial mRNA was PCR-amplified across the splice junction between exons 5 and 15. Transcripts corresponding to the hyaluronate receptor CD44H as well as a larger isoform were identified. CD44H was absent, or very scarce, in cDNA from purified gland epithelium. In contrast, Ishikawa cells expressed this form abundantly. The glands and Ishikawa cells also expressed CD44E containing sequences encoded by exons 12, 13, and 14. These data demonstrate the presence of CD44 in human endometrium and decidua, and show that different isoforms of CD44 are associated with tissue compartments in which different functional roles can be anticipated.

摘要

利用针对该分子所有形式共有的表位的六种单克隆抗体,通过免疫荧光法以及逆转录-聚合酶链反应(RT-PCR),检测了细胞表面黏附糖蛋白CD44在人子宫内膜中的分布情况。在整个月经周期的基质、血管、腺上皮和腔上皮中均观察到免疫反应性。观察到染色强度存在变化,尤其是在上皮区室。妊娠头三个月的蜕膜化基质细胞也强烈表达CD44。在正常周期各阶段未观察到免疫反应性的系统性变化,但有一部分(25%)标本的上皮缺乏反应性。为了确定上皮同种型的分子大小,开发了一种使用表面放射性碘化、去污剂提取的腺体的免疫沉淀技术。这表明在细胞表面存在一种单一的主要CD44E物种,其近似分子量为130 kDa。RT-PCR用于研究全子宫内膜组织、分离的腺体片段和石川子宫内膜癌细胞中存在的同种型。从总子宫内膜mRNA产生的互补DNA在第5外显子和第15外显子之间的剪接连接处进行PCR扩增。鉴定出了与透明质酸受体CD44H以及一种更大的同种型相对应的转录本。在纯化的腺上皮的cDNA中,CD44H不存在或非常稀少。相反,石川细胞大量表达这种形式。腺体和石川细胞还表达包含由第12、13和14外显子编码序列的CD44E。这些数据证明了CD44在人子宫内膜和蜕膜中的存在,并表明CD44的不同同种型与可以预期其具有不同功能作用的组织区室相关。

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