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Sample handling of gastric tissue and O-glycan alterations in paired gastric cancer and non-tumorigenic tissues

机译:配对胃癌和非致瘤组织中胃组织的样品处理和O-聚糖变化

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摘要

Sample collection, handling and storage are the most critical steps for ensuring the highest preservation of specimens. Pre-analytical variability can influence the results as protein signatures alter rapidly after tissue excision or during long-term storage. Hence, we evaluated current state-of-the-art biobank preservation methods from a glycomics perspective and analyzed O-glycan alterations occurring in the gastric cancer tissues. Paired tumor and adjacent normal tissue samples were obtained from six patients undergoing gastric cancer surgery. Collected samples (n = 24) were either snap-frozen or heat stabilized and then homogenized. Glycans were released from extracted glycoproteins and analyzed by LC-MS/MS. In total, the relative abundance of 83 O-glycans and 17 derived structural features were used for comparison. There was no statistically significant difference found in variables between snap frozen and heat-stabilized samples, which indicated the two preservation methods were comparable. The data also showed significant changes between normal and cancerous tissue. In addition to a shift from high sialylation in the cancer area towards blood group ABO in the normal area, we also detected that the LacdiNAc epitope (N,N’-diacetyllactosamine) was significantly decreased in cancer samples. The O-glycan alterations that are presented here may provide predictive power for the detection and prognosis of gastric cancer.
机译:样品的收集,处理和存储是确保最大程度保存标本的最关键步骤。分析前的可变性会影响结果,因为组织切除后或长期保存后蛋白质特征会迅速改变。因此,我们从糖组学的角度评估了当前最先进的生物库保存方法,并分析了在胃癌组织中发生的O型聚糖变化。从六名接受胃癌手术的患者中获得配对的肿瘤和邻近的正常组织样本。将收集的样品(n = 24)快速冷冻或热稳定,然后均质化。糖从提取的糖蛋白中释放出来,并通过LC-MS / MS分析。总共使用83个O-聚糖的相对丰度和17个衍生的结构特征进行比较。速冻样品和热稳定样品之间的变量之间没有统计学显着差异,这表明两种保存方法具有可比性。数据还显示正常组织和癌变组织之间有显着变化。除了从癌症区域的高唾液酸化转变为正常区域的ABO血型外,我们还检测到LacdiNAc表位(N,N'-二乙酰基乳糖胺)在癌症样本中显着降低。本文介绍的O-聚糖变化可能为胃癌的检测和预后提供预测力。

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