Expression of Podocalyxin Potentially Decorated With Low-sulfated Keratan Sulfate in Human Testicular Embryonal Carcinoma.

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Accounts of Chemical Research Pub Date : 2024-07-01 Epub Date: 2024-07-25 DOI:10.1369/00221554241265162

Akifumi Muramoto, Hitomi Hoshino, So Inamura, Masataka Murahashi, Tomoya O Akama, Naoki Terada, Motohiro Kobayashi

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Abstract

SummaryWe previously demonstrated that among various histological types of human testicular germinal cell tumors (GCTs), embryonal carcinoma (EC) preferentially expresses low-sulfated keratan sulfate (KS) consisting of repeating N-acetyllactosamine (LacNAc) disaccharide units composed of galactose and 6-O-sulfated N-acetylglucosamine (GlcNAc), which is recognized by the R-10G antibody. Recently, we generated another anti-low-sulfated KS monoclonal antibody, 294-1B1. Immunohistochemical analysis of testicular GCTs (n=83) revealed that the low-sulfated KS recognized by 294-1B1 is also preferentially expressed in EC but minimally in other GCT histological types. Moreover, immunolabeling with R-10G and 294-1B1 antibodies was resistant to peptide-N-glycosidase F digestion, and EC was not stained with the MECA-79 antibody, indicating that low-sulfated KS expressed in EC contains mucin-type core 2 O-glycans carrying GlcNAc-6-O-sulfated oligo-LacNAc. Double immunofluorescence staining showed that R-10G and 294-1B1 antibody signals colocalized with those for podocalyxin (PODXL). Furthermore, western blot analysis of recombinant human PODXL•IgG fusion proteins secreted from low-sulfated KS-expressing human embryonic kidney 293T cells revealed that PODXL functions as a core protein for low-sulfated KS. Taken together, these findings strongly suggest that the PODXL glycoform decorated with low-sulfated KS is preferentially expressed in human testicular EC and may therefore serve as a diagnostic marker for this malignancy.

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人睾丸胚胎癌中可能装饰有低硫酸化角叉菜胶的荚膜萼蛋白的表达

摘要我们曾证实，在各种组织学类型的人类睾丸生殖细胞瘤（GCTs）中，胚胎癌（EC）优先表达低硫酸化角蛋白硫酸盐（KS），该KS由重复的N-乙酰半乳糖胺（LacNAc）二糖单位组成，由半乳糖和6-O-硫酸化N-乙酰葡萄糖胺（GlcNAc）构成，可被R-10G抗体识别。最近，我们生成了另一种抗低硫酸化 KS 单克隆抗体 294-1B1，对睾丸 GCTs（n=83）进行免疫组化分析后发现，294-1B1 识别的低硫酸化 KS 也优先在 EC 中表达，但在其他 GCT 组织学类型中表达极少。此外，R-10G和294-1B1抗体的免疫标记对肽-N-糖苷酶F消化具有抗性，而MECA-79抗体不能对EC染色，这表明在EC中表达的低硫酸化KS含有携带GlcNAc-6-O-硫酸化寡糖-LacNAc的粘蛋白型核心2 O-聚糖。双重免疫荧光染色显示，R-10G 和 294-1B1 抗体信号与荚膜萼蛋白（PODXL）的信号共定位。此外，对表达低硫酸化 KS 的人胚肾 293T 细胞分泌的重组人 PODXL-IgG 融合蛋白进行的 Western 印迹分析表明，PODXL 是低硫酸化 KS 的核心蛋白。综上所述，这些发现有力地表明，用低硫酸化KS装饰的PODXL糖形在人类睾丸EC中优先表达，因此可作为这种恶性肿瘤的诊断标志物。

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.