肝内胆管癌（iCCA）是一种致命的恶性肿瘤，在全球范围内的发病率不断增加，治疗选择有限。异常蛋白质糖基化是癌症的主要特征之一。本研究深入探讨了富含脱氧葡萄糖元的糖基化在胆管癌发生中的可能作用。我们发现与非肿瘤邻近肝脏和正常胆管细胞相比，人iCCA组织中总脱氧葡萄糖元水平和富含脱氧葡萄糖元通路成员的表达普遍上调。此外，总脱氧葡萄糖元水平与患者预后不良相关。此外，使用6-炔基葡萄糖（6AF）抑制富含脱氧葡萄糖元后，iCCA细胞系的增殖和迁移呈剂量依赖性下降，而在细胞培养基中添加葡萄糖则会抵消这些影响。在分子水平上，FX合成酶和GDP-葡萄糖转膜转运体（SLC35C1）是细胞富含脱氧葡萄糖元的关键成分，通过FX/SLC35C1敲除或6AF处理可降低iCCA细胞系中的NOTCH活性、NOTCH1/Jagged1相互作用、NOTCH受体及相关靶基因。同时，在这些细胞中，EGFR、NF-κB p65和Bcl-xL蛋白水平减少，而IκBα（关键的细胞NF-kB抑制剂）在FX/SLC35C1敲除或6AF处理后增加。在鸟类胚胎绒毛尿囊膜（CAM）实验中，6AF处理大大抑制了iCCA细胞的生长。结论：升高的总脱氧葡萄糖元特征化了人类iCCA，通过上调NOTCH和EGFR/NF-kB途径促进细胞生长和迁移。因此，异常富含脱氧葡萄糖元是一种新的病理学参与者，也是人类iCCA的潜在治疗靶点。Copyright © 2023美国肝脏病研究协会。

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. We discovered that levels of global fucosylation and members of the fucosylation pathway are ubiquitously upregulated in human iCCA tissues compared to non-tumorous 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, NF-κB p65, and Bcl-xL protein levels diminished, whereas IκBα (a critical cellular NF-kB inhibitor) increased after FX/SLC35C1 knockdown or 6AF administration. In the chick chorioallantoic membrane (CAM) assay, 6AF treatment profoundly suppresses the growth of iCCA cells. Conclusions : Elevated global fucosylation characterizes human iCCA, contributing to cell growth and migration via the upregulation of the NOTCH and EGFR/NF-kB pathways. Thus, aberrant fucosylation is a novel pathogenetic player and a potential therapeutic target for human iCCA.Copyright © 2023 American Association for the Study of Liver Diseases.