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

Journal article
Authors Barbara Adamczyk
Chunsheng Jin
K. Polom
P. Munoz
Miguel A. Rojas-Macias
D. Zeeberg
M. Boren
F. Roviello
Niclas G. Karlsson
Published in Scientific Reports
Volume 8
ISSN 2045-2322
Publication year 2018
Published at Institute of Biomedicine, Department of Medical Biochemistry and Cell Biology
Language en
Keywords mass-spectrometry, glycoprofiling platform, linked oligosaccharides, glycosylation analysis, unicarb-db, extraction, stabilization, glycoproteins, nomenclature, biomarker
Subject categories Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy), Cancer and Oncology


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.

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