Duran Adrià, Guerrero Pedro E, Ortiz Maria Rosa, Pérez Del Campo Dúnia, Castro Ernesto, Garcia-Velasco Adelaida, Fort Esther, de Llorens Rafael, Saldova Radka, Llop Esther, Peracaula Rosa
Biochemistry and Molecular Biology Unit, Department of Biology, University of Girona, 17003 Girona, Spain.
Girona Biomedical Research Institute (IDIBGI), 17190 Girona, Spain.
Biomedicines. 2022 Aug 10;10(8):1942. doi: 10.3390/biomedicines10081942.
Currently, there are no reliable biomarkers for the diagnosis of pancreatic cancer (PaC). Glycoproteomic approaches that analyze the glycan determinants on specific glycoproteins have proven useful to develop more specific cancer biomarkers than the corresponding protein levels. In PaC, mesothelin (MSLN) is a neo-expressed glycoprotein. MSLN glycosylation has not been described and could be altered in PaC. In this work, we aimed to characterize MSLN glycans from PaC cells and serum samples to assess their potential usefulness as PaC biomarkers. First, we analyzed MSLN glycans from PaC cell lines and then we developed an enzyme-linked lectin assay to measure core fucosylated-MSLN (Cf-MSLN) glycoforms. MSLN glycans from PaC cells were analyzed by glycan sequencing and through Western blotting with lectins. All of the cell lines secreted MSLN, with its three -glycosylation sites occupied by complex-type -glycans, which were mainly α2,3-sialylated, core fucosylated and highly branched. The Cf-MSLN glycoforms were quantified on PaC serum samples, and compared with MSLN protein levels. The Cf-MSLN was significantly decreased in PaC patients compared to control sera, while no differences were detected by using MSLN protein levels. In conclusion, Cf-MSLN glycoforms were differently expressed in PaC, which opens the way to further investigate their usefulness as PaC biomarkers.
目前，尚无可靠的生物标志物用于胰腺癌（PaC）的诊断。与相应的蛋白质水平相比，分析特定糖蛋白上聚糖决定簇的糖蛋白质组学方法已被证明有助于开发更具特异性的癌症生物标志物。在胰腺癌中，间皮素（MSLN）是一种新表达的糖蛋白。尚未对MSLN糖基化进行描述，并且在胰腺癌中可能会发生改变。在这项研究中，我们旨在表征来自胰腺癌细胞和血清样本的MSLN聚糖，以评估它们作为胰腺癌生物标志物的潜在用途。首先，我们分析了胰腺癌细胞系中的MSLN聚糖，然后开发了一种酶联凝集素测定法来测量核心岩藻糖基化的MSLN（Cf-MSLN）糖型。通过聚糖测序和使用凝集素的蛋白质印迹法分析胰腺癌细胞中的MSLN聚糖。所有细胞系均分泌MSLN，其三个糖基化位点被复合型聚糖占据主要是α2,3-唾液酸化、核心岩藻糖基化且高度分支。对胰腺癌血清样本中的Cf-MSLN糖型进行定量，并与MSLN蛋白质水平进行比较。与对照血清相比，胰腺癌患者的Cf-MSLN显著降低，而使用MSLN蛋白质水平未检测到差异。总之，Cf-MSLN糖型在胰腺癌中表达不同，这为进一步研究它们作为胰腺癌生物标志物的用途开辟了道路。
