Atatürk University, Faculty of Pharmacy, Department of Analytical Chemistry, Erzurum, 25240, Turkey.
Mol Omics. 2021 Jun 14;17(3):394-404. doi: 10.1039/d0mo00150c.
Invasive ductal carcinoma (IDC) is the most common type of breast cancer. As dynamic changes of the glycome are closely associated with complex diseases, they have become a focal point of cancer research involving predictive and prognostic markers. Formalin-fixed paraffin-embedded (FFPE) clinical specimens are representative of the tumor environment and are thus utilized in studies on cancer related research and biomarker discovery. Further studies on differential N-glycosylation profiling of IDC cancer tissues are necessary in order to understand the biological role of glycans in cancer and to evaluate their predictive ability. In this study, matrix assisted laser desorption ionization-mass spectrometry (MALDI-MS)-based analyses were conducted for determining differential N-glycosylation patterns of IDC. Two different derivatization methods, namely, 2-aminobenzoic acid (2-AA) labeling and linkage-specific sialic acid esterification, were used for the analysis of N-glycans. Forty-seven 2-AA labeled and fifty ethyl esterified N-glycans were identified by MALDI-MS. In statistical analyses conducted for 2-AA-labeled N-glycans, the relative amounts of 32 N-glycans and prevalence of 15 N-glycan traits showed significant (p < 0.05) differences between cancer and normal tissues; and in such analyses for the ethyl-esterified N-glycans, the relative amounts of 27 N-glycans and prevalence of 17 N-glycan traits showed significant (p < 0.05) differences between them. It was found that mainly high mannose N-glycans, including H5N2, H6N2, and H7N2, and two fucosylated compositions (H3N3F1 and H5N5F1) showed strong discrimination between IDC and controls. In addition, compared with the controls, high mannose N-glycans were observed to be up-regulated in IDC whereas bisecting N-glycans were down-regulated.
浸润性导管癌（IDC）是最常见的乳腺癌类型。由于聚糖的动态变化与复杂疾病密切相关，因此它们已成为涉及预测和预后标志物的癌症研究的焦点。福尔马林固定石蜡包埋（FFPE）临床标本代表肿瘤环境，因此可用于癌症相关研究和生物标志物发现的研究。为了了解糖在癌症中的生物学作用并评估其预测能力，有必要进一步研究 IDC 癌组织中差异的 N-糖基化谱。在这项研究中，进行了基质辅助激光解吸电离 - 质谱（MALDI-MS）分析，以确定 IDC 的差异 N-糖基化模式。使用了两种不同的衍生化方法，即 2-氨基苯甲酸（2-AA）标记和连接特异性唾液酸酯化，用于 N-聚糖的分析。通过 MALDI-MS 鉴定了 47 个 2-AA 标记和 50 个乙酯化 N-聚糖。在对 2-AA 标记的 N-聚糖进行的统计分析中，32 个 N-聚糖的相对量和 15 个 N-聚糖特征的出现率在癌症和正常组织之间表现出显著差异（p <0.05）；在对乙酯化 N-聚糖进行的此类分析中，27 个 N-聚糖的相对量和 17 个 N-聚糖特征的出现率在癌症和正常组织之间表现出显著差异（p <0.05）。结果发现，主要是高甘露糖 N-聚糖，包括 H5N2、H6N2 和 H7N2，以及两种岩藻糖组成（H3N3F1 和 H5N5F1），在 IDC 和对照组之间表现出强烈的区分。此外，与对照组相比，在 IDC 中观察到高甘露糖 N-聚糖上调，而双分支 N-聚糖下调。
