Inflammatory cytokines regulate the expression of glycosyltransferases involved in the biosynthesis of tumor-associated sialylated glycans in pancreatic cancer cell lines.
BACKGROUND
Pancreatic ductal adenocarcinoma (PDAC) is characterized by an abundant stroma containing several pro-inflammatory cytokines, which are described to modulate the expression of important genes related to tumor promotion and progression. In the present work we have investigated the potential role of these cytokines in the biosynthesis of tumor-associated carbohydrate antigens such as sialyl-Lewis(x) (SLe(x)) through the regulation of specific glycosyltransferase genes.


METHODS
Two human PDAC cell lines MDAPanc-3 and MDAPanc-28 were treated with pro-inflammatory cytokines IL-1β, TNFα, IL-6 or IL-8, and the content of tumor-associated carbohydrate antigens at the cell membrane was analyzed by flow cytometry. In addition, variation in the mRNA expression of sialyltransferase (ST) and fucosyltransferase (FUT) genes, which codify for the ST and FucT enzymes involved in the carbohydrate antigens' biosynthesis, was determined. The inflammatory microenvironment of PDAC tissues and the expression of Lewis-type antigens were analyzed by immunohistochemistry to find a possible correlation between inflammation status and the presence of tumor-associated carbohydrate antigens.


RESULTS
IL-1β stimuli increased SLe(x) and α2,6-sialic acid levels in MDAPanc-28 cells and enhanced the mRNA levels of ST3GAL3-4 and FUT5-7, which codify for ST and FucT enzymes related to SLe(x) biosynthesis, and of ST6GAL1. IL-6 and TNFα treatments increased the levels of SLe(x) and Le(y) antigens in MDPanc-3 cells and, similarly, the mRNA expression of ST3GAL3-4, FUT1-2 and FUT6, related to these Lewis-type antigens' biosynthesis, were increased. Most PDAC tissues stained for SLe(x) and SLe(a) and tended to be expressed in the tumor samples with a higher presence of inflammatory immune cells.


CONCLUSIONS
The inflammatory microenvironment can modulate the glycosylation pattern of PDAC cells, increasing the expression of tumor-associated sialylated antigens such as SLe(x), which contributes to pancreatic tumor malignancy.
Biochemistry and Molecular Biology Unit, Department of Biology, University of Girona, Girona, Spain.
Sònia
S
Bassagañas
Department of Pathology, Vall d'Hebron University Hospital, Barcelona, Spain.
Helena
H
Allende
Institut Hospital del Mar d'Investigacions Mèdiques (IMIM), Barcelona, Spain.
Lara
L
Cobler
Department of Pathology, Dr. Josep Trueta University Hospital, Girona, Spain.
M Rosa
MR
Ortiz
Biochemistry and Molecular Biology Unit, Department of Biology, University of Girona, Girona, Spain.
Esther
E
Llop
Department of Pathology, Vall d'Hebron University Hospital, Barcelona, Spain.
Carme
C
de Bolós
Biochemistry and Molecular Biology Unit, Department of Biology, University of Girona, Girona, Spain. Electronic address: rosa.peracaula@udg.edu.
Rosa
R
Peracaula
D021441:Carcinoma, Pancreatic Ductal / Q000378:metabolism*; D045744:Cell Line, Tumor; D016207:Cytokines / Q000378:metabolism*; D018450:Disease Progression; D000939:Epitopes / Q000737:chemistry; D005434:Flow Cytometry; D015971:Gene Expression Regulation, Enzymologic; D015972:Gene Expression Regulation, Neoplastic; D016695:Glycosyltransferases / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D007249:Inflammation / Q000378:metabolism*; D053583:Interleukin-1beta / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D016256:Lewis X Antigen / Q000737:chemistry; D009844:Oligosaccharides / Q000378:metabolism; D010190:Pancreatic Neoplasms / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism*; D012794:Sialic Acids / Q000737:chemistry; D000080506:Sialyl Lewis X Antigen; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D016207:Cytokines; D000939:Epitopes; D053583:Interleukin-1beta; D015850:Interleukin-6; D016209:Interleukin-8; D016256:Lewis X Antigen; D009844:Oligosaccharides; D011134:Polysaccharides; D012794:Sialic Acids; D000080506:Sialyl Lewis X Antigen; D014409:Tumor Necrosis Factor-alpha; D016695:Glycosyltransferases
Cytokines; Fucosyltransferases; Lewis antigens; Pancreatic adenocarcinoma; Sialyltransferases