Inhibition of Fucosylation in Human Invasive Ductal Carcinoma Reduces E-Selectin Ligand Expression, Cell Proliferation, and ERK1/2 and p38 MAPK Activation

• Main Author: Carrascal, M
• Publication Date: 2018
• Other Authors: Silva, M, Ramalho, J, Pen, C, Martins, M, Pascoal, C, Amaral, C, Serrano, I, Oliveira, MJ, Sackstein, R, Videira, P
• Format: Article
• Language: eng
• Source: Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
• Download full: http://hdl.handle.net/10400.17/3949
• topic: HSJ ANPAT; Adult; Aged; Female; Humans; Breast Neoplasms / enzymology*; Middle Aged; Breast Neoplasms / pathology; Carcinoma, Ductal, Breast / enzymology*; Carcinoma, Ductal, Breast / pathology; Cell Adhesion / drug effects; Cell Line, Tumor; Cell Proliferation / drug effects; E-Selectin / genetics; E-Selectin / metabolism*; Fucose / analogs & derivatives; Fucose / pharmacology; Fucosyltransferases / antagonists & inhibitors*; Ligands; MAP Kinase Signaling System / drug effects; Neoplasm Invasiveness; Oligosaccharides / metabolism*; Primary Cell Culture; Sialyl Lewis X Antigen; p38 Mitogen-Activated Protein Kinases / genetics; p38 Mitogen-Activated Protein Kinases / metabolism*

Breast cancer tissue overexpresses fucosylated glycans, such as sialyl-Lewis X/A (sLeX/A ), and α-1,3/4-fucosyltransferases (FUTs) in relation to increased disease progression and metastasis. These glycans in tumor circulating cells mediate binding to vascular E-selectin, initiating tumor extravasation. However, their role in breast carcinogenesis is still unknown. Here, we aimed to define the contribution of the fucosylated structures, including sLeX/A , to cell adhesion, cell signaling, and cell proliferation in invasive ductal carcinomas (IDC), the most frequent type of breast cancer. We first analyzed expression of E-selectin ligands in IDC tissue and established primary cell cultures from the tissue. We observed strong reactivity with E-selectin and anti-sLeX/A antibodies in both IDC tissue and cell lines, and expression of α-1,3/4 FUTs FUT4, FUT5, FUT6, FUT10, and FUT11. To further assess the role of fucosylation in IDC biology, we immortalized a primary IDC cell line with human telomerase reverse transcriptase to create the 'CF1_T cell line'. Treatment with 2-fluorofucose (2-FF), a fucosylation inhibitor, completely abrogated its sLeX/A expression and dramatically reduced adherence of CF1_T cells to E-selectin under hemodynamic flow conditions. In addition, 2-FF-treated CF1_T cells showed a reduced migratory ability, as well as decreased cell proliferation rate. Notably, 2-FF treatment lowered the growth factor expression of CF1_T cells, prominently for FGF2, vascular endothelial growth factor, and transforming growth factor beta, and negatively affected activation of signal-regulating protein kinases 1 and 2 and p38 mitogen-activated protein kinase signaling pathways. These data indicate that fucosylation licenses several malignant features of IDC, such as cell adhesion, migration, proliferation, and growth factor expression, contributing to tumor progression.