Abstract
BackgroundSeveral reports have shown the role of glycosylation in pancreatic cancer (PC), but a global systematic screening of specific glycosyltransferases (glycoTs) in its progression remains unknown.MethodsWe demonstrate a rigorous top-down approach using TCGA-based RNA-Seq analysis, multi-step validation using RT-qPCR, immunoblots and immunohistochemistry. We identified six unique glycoTs (B3GNT3, B4GALNT3, FUT3, FUT6, GCNT3 and MGAT3) in PC pathogenesis and studied their function using CRISPR/Cas9-based KD systems.ResultsSerial metastatic in vitro models using T3M4 and HPAF/CD18, generated in house, exhibited decreases in B3GNT3, FUT3 and GCNT3 expression on increasing metastatic potential. Immunohistochemistry identified clinical significance for GCNT3, B4GALNT3 and MGAT3 in PC. Furthermore, the effects of B3GNT3, FUT3, GCNT3 and MGAT3 were shown on proliferation, migration, EMT and stem cell markers in CD18 cell line. Talniflumate, GCNT3 inhibitor, reduced colony formation and migration in T3M4 and CD18 cells. Moreover, we found that loss of GCNT3 suppresses PC progression and metastasis by downregulating cell cycle genes and β-catenin/MUC4 axis. For GCNT3, proteomics revealed downregulation of MUC5AC, MUC1, MUC5B including many other proteins.ConclusionsCollectively, we demonstrate a critical role of O- and N-linked glycoTs in PC progression and delineate the mechanism encompassing the role of GCNT3 in PC.
Highlights
Several reports have shown the role of glycosylation in pancreatic cancer (PC), but a global systematic screening of specific glycosyltransferases in its progression remains unknown
Top-down transcriptomic analysis reveals overexpression of B3GNT3, B4GALNT3, FUT3, FUT6, GCNT3 and MGAT3 in PC There are more than 200 glycosyltransferases that facilitate the enzymatic addition of a variety of carbohydrate structures in the human cell.[21]
GlycoTs genes overexpressed are as follows: A4GNT, ABO, ALG1L, B3GALT5, B3GNT3, B3GNT5, B3GNT6, B3GNT7, B4GALNT2, B4GALNT3, B4GALT4, C1GALT1, CHPF, COLGALT2, FUT2, FUT3, FUT6, FUT9, GALNT12, GALNT3, GALNT5,GCNT3, HAS1, HAS2, HAS3, LFNG, MGAT3, PYGB, ST6GALNAC1, ST8SIA2, UGT1A6, UGT1A7, UGT1A8, UGT2B4, UGT2B7 and UGT8
Summary
Several reports have shown the role of glycosylation in pancreatic cancer (PC), but a global systematic screening of specific glycosyltransferases (glycoTs) in its progression remains unknown. We identified six unique glycoTs (B3GNT3, B4GALNT3, FUT3, FUT6, GCNT3 and MGAT3) in PC pathogenesis and studied their function using CRISPR/Cas9-based KD systems. The discovery of CA19-9 as a diagnostic marker for PC has helped to address some of the challenges.[3,4,5] CA19-9 is a sLea-type glycan structure found either on cell membranes or on circulating secretory glycoproteins. The discovery of this marker towards the diagnosis of PC has helped in understanding the importance of glycogenes in PC.
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