High-Throughput Mass Spectrometry Analysis of N-Glycans and Protein Markers after FUT8 Knockdown in the Syngeneic SW480/SW620 Colorectal Cancer Cell Model,Journal of Proteome Research

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High-Throughput Mass Spectrometry Analysis of N-Glycans and Protein Markers after FUT8 Knockdown in the Syngeneic SW480/SW620 Colorectal Cancer Cell Model
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2024-03-20 , DOI: 10.1021/acs.jproteome.3c00833
Rubén López-Cortés ₁ , Laura Muinelo-Romay ₂ , Almudena Fernández-Briera ₃ , Emilio Gil Martín ₄

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Disruption of the glycosylation machinery is a common feature in many types of cancer, and colorectal cancer (CRC) is no exception. Core fucosylation is mediated by the enzyme fucosyltransferase 8 (FucT-8), which catalyzes the addition of α1,6-l-fucose to the innermost GlcNAc residue of N-glycans. We and others have documented the involvement of FucT-8 and core-fucosylated proteins in CRC progression, in which we addressed core fucosylation in the syngeneic CRC model formed by SW480 and SW620 tumor cell lines from the perspective of alterations in their N-glycosylation profile and protein expression as an effect of the knockdown of the FUT8 gene that encodes FucT-8. Using label-free, semiquantitative mass spectrometry (MS) analysis, we found noticeable differences in N-glycosylation patterns in FUT8-knockdown cells, affecting core fucosylation and sialylation, the Hex/HexNAc ratio, and antennarity. Furthermore, stable isotopic labeling of amino acids in cell culture (SILAC)-based proteomic screening detected the alteration of species involved in protein folding, endoplasmic reticulum (ER) and Golgi post-translational stabilization, epithelial polarity, and cellular response to damage and therapy. This data is available via ProteomeXchange with identifier PXD050012. Overall, the results obtained merit further investigation to validate their feasibility as biomarkers of progression and malignization in CRC, as well as their potential usefulness in clinical practice.

中文翻译：

同基因 SW480/SW620 结直肠癌细胞模型中 FUT8 敲除后 N-聚糖和蛋白质标记物的高通量质谱分析

糖基化机制的破坏是许多类型癌症的共同特征，结直肠癌 (CRC) 也不例外。核心岩藻糖基化由岩藻糖基转移酶 8 (FucT-8) 介导，该酶催化将 α1,6- l-岩藻糖添加到N -聚糖最内部的 GlcNAc 残基上。我们和其他人已经记录了FucT-8和核心岩藻糖基化蛋白在CRC进展中的参与，其中我们从SW480和SW620肿瘤细胞系形成的同基因CRC模型中从N-糖基化谱变化的角度解决了核心岩藻糖基化问题编码 FucT-8 的FUT8基因敲低后的蛋白质表达。使用无标记、半定量质谱 (MS) 分析，我们发现FUT8敲低细胞中N -糖基化模式存在显着差异，影响核心岩藻糖基化和唾液酸化、Hex/HexNAc比率和触角。此外，基于细胞培养中氨基酸的稳定同位素标记 (SILAC) 的蛋白质组学筛选检测到参与蛋白质折叠、内质网 (ER) 和高尔基体翻译后稳定、上皮极性以及细胞对损伤和治疗的反应的物种的改变。该数据可通过 ProteomeXchange 获得，标识符为 PXD050012。总的来说，这些结果值得进一步研究，以验证它们作为结直肠癌进展和恶性化生物标志物的可行性，以及它们在临床实践中的潜在用途。

更新日期：2024-03-20

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