N-acetylglucosaminyltransferase V negatively regulates integrin α5β1-mediated monocyte adhesion and transmigration through vascular endothelium

Yang,Huo-Mei; Yu,Chao; Yang,Zhu

doi:10.3892/ijo.2012.1484

N-acetylglucosaminyltransferase V negatively regulates integrin α5β1-mediated monocyte adhesion and transmigration through vascular endothelium

Authors:
- Huo-Mei Yang
- Chao Yu
- Zhu Yang
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Affiliations: Department of Obstetrics and Gynecology, the Second Affiliated Hospital of Chongqing Medical University, Chongqing, Chongqing 400010, P.R. China, Institute of Life Sciences, Chongqing Medical University, Chongqing, Chongqing 400016, P.R. China
Published online on: May 17, 2012 https://doi.org/10.3892/ijo.2012.1484
Pages: 589-598

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Abstract

Changes in the expression of glycosyltransferases that branch N-linked glycans are associated with many physiological and pathological events, such as cell adhesion, migration, proliferation and tumor cell malignancy. Here, the altered levels of N-acetylglucosaminyltransferase V (GnT-V) and its product β(1,6)-linked GlcNAc in monocytes were observed during inflammation. The effects of GnT-V and aberrant N-linked β(1,6) branching on monocyte adhesion through vascular endothelium and transmigration were investigated. During IFN-γ-induced inflammation, adhesion and transendothelial migration of THP-1 monocytes were enhanced, and the levels of GnT-V and β(1,6)-linked GlcNAc in THP-1 monocytes were significantly decreased. Expression of the GnT-V shRNA vector in THP-1 cells reversed the abnormal IFN-γ-induced characteristics, indicating direct involvement of N-glycosylation in these biological effects. The enhanced adhesion and transendothelial migration were significantly inhibited by functional blockade with antibodies against integrin α5 or β1 in IFN-γ-induced and GnT-V knockdown THP-1 cells, demonstrating the involvement of integrin α5β1 in the monocyte-endothelium interaction. However, IFN-γ treatment and GnT-V knockdown in THP-1 cells lowered expression of N-linked β(1,6) branching on integrin α5 and β1, without affecting the total protein expression of the subunits. Decreased GnT-V expression caused marked enchancement of integrin-induced phosphorylation of focal adhesion kinase (FAK). The augmented FAK-mediated ERK phosphorylation and activation were observed in IFN-γ-induced THP-1 cells. Furthermore, ERK inhibitor pre-treatment nearly abrogated the highly elevated IFN-γ-induced monocyte adhesion and transmigration, concomitant with reversal of the decrease in GnT-V and β(1,6) branching. Our results demonstrate for the first time that decreased GnT-V activity due to inflammatory cytokine induction in human monocytes resulted in enchancement of integrin α5β1-dependent monocyte-vascular endothelium adhesion and transmigration. Consequently, the activation of integrin caused elevation of FAK phosphorylation. These effects promoted FAK-mediated downstream signaling, including the ERK pathway, and indicate that GnT-V may be a potential therapeutic target for vascular inflammatory conditions.

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