Barrabés Sílvia, Pagès-Pons Lluís, Radcliffe Catherine M, Tabarés Glòria, Fort Esther, Royle Louise, Harvey David J, Moenner Michel, Dwek Raymond A, Rudd Pauline M, De Llorens Rafael, Peracaula Rosa
Unitat de Bioquímica i Biologia Molecular, Departament de Biologia, Universitat de Girona, Campus de Montilivi, Girona 17071, Spain.
Glycobiology. 2007 Apr;17(4):388-400. doi: 10.1093/glycob/cwm002. Epub 2007 Jan 17.
Human pancreatic ribonuclease 1 (RNase 1) is a glycoprotein expressed mainly by the pancreas and also found in endothelial cells. The diagnosis of pancreatic cancer (PaC) remains difficult and therefore the search for sensitive and specific markers is required. Previous studies showed that RNase 1 from human healthy pancreas contained only neutral glycans, whereas RNase 1 from PaC cell lines contained sialylated structures. To determine whether these glycan tumor cell-associated changes were also characteristic of serum RNase 1 and could be used as a marker of PaC, we have analyzed the glycosylation of serum RNase 1. The origin of serum RNase 1 was also investigated. Serum RNase 1 from two PaC patients and two controls was purified and the glycans analyzed by high-performance liquid chromatography (HPLC)-based sequencing and mass spectrometry. Although normal and tumor serum RNase 1 contained the same glycan structures, there was an increase of 40% in core fucosylation in the main sialylated biantennary glycans in the PaC serum RNase 1. This change in proportion would be indicative of a subset of tumor-associated glycoforms of RNase 1, which may provide a biomarker for PaC. Two-dimensional electrophoresis of the RNase 1 from several endothelial cell lines, EA.hy926, human umbilical vein endothelial cells (HUVEC), human mammary microvessel endothelial cells (HuMMEC), and human lung microvessel endothelial cells (HuLEC), showed basically the same pattern and was also very similar to that of serum RNase 1. RNase 1 from EA.hy926 was then purified and presented a glycosylation profile very similar to that from serum RNase 1, suggesting that endothelial cells are the main source of this enzyme.
人胰腺核糖核酸酶1（RNase 1）是一种主要由胰腺表达的糖蛋白，在内皮细胞中也有发现。胰腺癌（PaC）的诊断仍然困难，因此需要寻找敏感且特异的标志物。先前的研究表明，来自健康人胰腺的RNase 1仅含有中性聚糖，而来自PaC细胞系的RNase 1含有唾液酸化结构。为了确定这些与肿瘤细胞相关的聚糖变化是否也是血清RNase 1的特征，以及是否可作为PaC的标志物，我们分析了血清RNase 1的糖基化情况。同时还研究了血清RNase 1的来源。从两名PaC患者和两名对照者中纯化血清RNase 1，并通过基于高效液相色谱（HPLC）的测序和质谱分析聚糖。尽管正常和肿瘤血清RNase 1含有相同的聚糖结构，但PaC血清RNase 1中主要唾液酸化双天线聚糖的核心岩藻糖基化增加了40%。这种比例变化表明RNase 1存在与肿瘤相关的糖型亚群，这可能为PaC提供一种生物标志物。对几种内皮细胞系EA.hy926、人脐静脉内皮细胞（HUVEC）、人乳腺微血管内皮细胞（HuMMEC）和人肺微血管内皮细胞（HuLEC）的RNase 1进行二维电泳，结果显示基本相同的模式，且与血清RNase 1的模式非常相似。然后纯化了EA.hy926细胞的RNase 1，其糖基化谱与血清RNase 1的非常相似，表明内皮细胞是这种酶的主要来源。
