Aberrant fucosylation sustains the NOTCH and EGFR/NF-κB pathways and has a prognostic value in human intrahepatic cholangiocarcinoma

Abstract

Background and Aims: Intrahepatic cholangiocarcinoma (iCCA) is a lethal malignancy, with increasing incidence worldwide and limited therapeutic options. Aberrant protein glycosylation is a hallmark of cancer. Here, we thoroughly investigated the possible involvement of fucosylation in cholangiocarcinogenesis. Approach and Results: We discovered that the levels of global fucosylation and members of the fucosylation pathway are ubiquitously upregulated in human iCCA tissues compared to nontumorous surrounding livers and normal biliary cells. In addition, total fucosylation levels correlate with poor patients’ prognosis. Furthermore, fucosylation inhibition following 6-alkynylfucose (6AF) administration triggered a dose-dependent decrease in the proliferation and migration of iCCA cell lines. Notably, adding fucose to the cell medium annulled these effects. At the molecular level, 6AF administration or small interfering RNA-mediated silencing of GDP-L-fucose synthetase (FX) and the GDP-fucose transmembrane transporter (SLC35C1), both pivotal players of cellular fucosylation, decreased NOTCH activity, NOTCH1/Jagged1 interaction, NOTCH receptors, and related target genes in iCCA cell lines. In the same cells, EGFR, nuclear factor kappa-light-chain-enhancer of activated B cells p65, and Bcl-xL protein levels diminished, whereas IκBα (a critical cellular NF-κB inhibitor) increased after FX/SLC35C1 knockdown or 6AF administration. In the chick chorioallantoic membrane assay, 6AF treatment profoundly suppresses the growth of iCCA cells. Conclusions: Elevated global fucosylation characterizes human iCCA, contributing to cell growth and migration through the upregulation of the NOTCH and EGFR/NF-κB pathways. Thus, aberrant fucosylation is a novel pathogenetic player and a potential therapeutic target for human iCCA.

O-GlcNAcylation of Keratin 18 coordinates TCA cycle to promote cholangiocarcinoma progression

“…34 Glyco-analytical methodologies including lectin blot, 35 glycan microarray 36 and glycoproteomic analysis 37,38 have been utilized to probe the glycobiology in CCA. In addition, abnormal sialylation, 39 fucosylation 40 and O-linked N-acetylgalactosamine (GalNAc) modi cation (O-GalNAcylation) 41 have also been implicated in prognostic outcome in human cholangiocarcinoma. Changes in O-GlcNAc levels and the cycling enzyme pair OGT/OGA in a myriad of tumor types have also been reviewed.…”

O-GlcNAcylation of Keratin 18 coordinates TCA cycle to promote cholangiocarcinoma progression

“…55,56 Glyco-analytical methodologies including lectin blot, [57][58][59] glycan microarray 60 and glycoproteomic analysis [61][62][63][64] have been utilized to probe the glycobiology in CCA. In addition, abnormal sialylation, [65][66][67][68] fucosylation 69,70 and O-linked Nacetylgalactosamine (GalNAc) modification (O-GalNAcylation) 60,[71][72][73] have also been implicated in prognostic outcome in human cholangiocarcinoma. Changes in O-GlcNAc levels and the cycling enzyme pair OGT/OGA in a myriad of tumor types have also been reviewed.…”

Notch signaling pathway in cancer: from mechanistic insights to targeted therapies

“… 118 Another study revealed that elevated fucosylation is a hallmark of human iCCA, promoting cell growth and migration by upregulating Notch and EGFR/NF-κB pathways. 119 Simultaneously, the Notch pathway is considered a key indicator of CCA progression and prognosis. 120 , 121 Studies show that Notch1 is upregulated in iCCA, potentially promoting iCCA migration by inducing EMT.…”