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人表皮糖蛋白/粘蛋白21(MUC21)的鉴定与表达:一种新型跨膜粘蛋白

Identification and expression of human epiglycanin/MUC21: a novel transmembrane mucin.

作者信息

Itoh Yuichi, Kamata-Sakurai Mika, Denda-Nagai Kaori, Nagai Shigenori, Tsuiji Makoto, Ishii-Schrade Katrin, Okada Kyoko, Goto Akiteru, Fukayama Masashi, Irimura Tatsuro

机构信息

Laboratory of Cancer Biology and Molecular Immunology, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113-0033, Japan.

出版信息

Glycobiology. 2008 Jan;18(1):74-83. doi: 10.1093/glycob/cwm118. Epub 2007 Oct 31.

Abstract

The gene for the human orthologue of mouse epiglycanin, a mucin expressed on mammary carcinoma TA3-Ha cells but not TA3-St cells, was identified by homology search to a mouse epiglycanin cDNA fragment identified by representational difference analysis between TA3-Ha and TA3-St cells. The open reading frame of this gene was cloned from human cervical carcinoma ME-180 cells. It consists of a mucin domain with 28 nonidentical tandem repeats of 45 nucleotides each corresponding to a threonine/serine-rich peptide, a stem domain, a transmembrane domain, and a cytoplasmic tail. The cloned cDNA with a FLAG sequence was expressed in K562 cells. A combination of immunoprecipitation with a polyclonal antibody specific for the cytoplasmic tail and Western blotting analysis with an anti-FLAG antibody and lectins revealed a mucin-like component as the gene product. Analysis by the use of tissue cDNA libraries indicated that the gene is expressed in lung, large intestine, thymus, and testis among 16 normal tissues tested. The polyclonal antibody specific for a synthetic peptide from the cytoplasmic tail, when tested for its reactivity with normal lung tissues, reacted with epithelia of bronchi and bronchioli but not with alveoli. All of 24 lung adenocarcinomas specimens tested were reactive with the antibody, whereas reactivity was observed with only 2 out of 24 squamous and none out of 24 small cell lung carcinomas. This is a novel transmembrane mucin and designated as MUC21.

摘要

小鼠表位糖蛋白的人类同源基因,该糖蛋白在乳腺癌TA3-Ha细胞而非TA3-St细胞上表达,通过与小鼠表位糖蛋白cDNA片段进行同源性搜索得以鉴定,该片段是通过TA3-Ha和TA3-St细胞之间的代表性差异分析确定的。该基因的开放阅读框是从人宫颈癌ME-180细胞中克隆出来的。它由一个粘蛋白结构域组成,该结构域有28个不相同的45个核苷酸的串联重复序列,每个重复序列对应一个富含苏氨酸/丝氨酸的肽段,一个茎结构域,一个跨膜结构域和一个细胞质尾巴。带有FLAG序列的克隆cDNA在K562细胞中表达。用针对细胞质尾巴的多克隆抗体进行免疫沉淀,并用抗FLAG抗体和凝集素进行蛋白质印迹分析,结果显示该基因产物是一种粘蛋白样成分。通过使用组织cDNA文库分析表明,在所检测的16种正常组织中,该基因在肺、大肠、胸腺和睾丸中表达。针对细胞质尾巴合成肽的多克隆抗体在检测其与正常肺组织的反应性时,与支气管和细支气管的上皮细胞发生反应,但不与肺泡发生反应。所检测的24例肺腺癌标本均与该抗体发生反应,而在24例鳞状细胞癌中只有2例有反应,在24例小细胞肺癌中均无反应。这是一种新型跨膜粘蛋白,并命名为MUC21。

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