Cell Surface Glycan Alterations in Epithelial Mesenchymal Transition Process of Huh7 Hepatocellular Carcinoma Cell

Abstract

Background and ObjectiveDue to recurrence and metastasis, the mortality of Hepatocellular carcinoma (HCC) is high. It is well known that the epithelial mesenchymal transition (EMT) and glycan of cell surface glycoproteins play pivotal roles in tumor metastasis. The goal of this study was to identify HCC metastasis related differential glycan pattern and their enzymatic basis using a HGF induced EMT model.MethodologyHGF was used to induce HCC EMT model. Lectin microarray was used to detect the expression of cell surface glycan and the difference was validated by lectin blot and fluorescence cell lectin-immunochemistry. The mRNA expression levels of glycotransferases were determined by qRT-PCR.ResultsAfter HGF treatment, the Huh7 cell lost epithelial characteristics and obtained mesenchymal markers. These changes demonstrated that HGF could induce a typical cell model of EMT. Lectin microarray analysis identified a decreased affinity in seven lectins ACL, BPL, JAC, MPL, PHA-E, SNA, and SBA to the glycan of cell surface glycoproteins. This implied that glycan containing T/Tn-antigen, NA2 and bisecting GlcNAc, Siaα2-6Gal/GalNAc, terminal α or βGalNAc structures were reduced. The binding ability of thirteen lectins, AAL, LCA, LTL, ConA, NML, NPL, DBA, HAL, PTL II, WFL, ECL, GSL II and PHA-L to glycan were elevated, and a definite indication that glycan containing terminal αFuc and ± Sia-Le, core fucose, α-man, gal-β(α) GalNAc, β1,6 GlcNAc branching and tetraantennary complex oligosaccharides structures were increased. These results were further validated by lectin blot and fluorescence cell lectin-immunochemistry. Furthermore, the mRNA expression level of Mgat3 decreased while that of Mgat5, FucT8 and β3GalT5 increased. Therefore, cell surface glycan alterations in the EMT process may coincide with the expression of glycosyltransferase.ConclusionsThe findings of this study systematically clarify the alterations of cell surface glycan in cancer EMT, and may provide novel insight for HCC metastasis.

β-Galactoside α2,6-Sialyltranferase 1 Promotes Transforming Growth Factor-β-mediated Epithelial-Mesenchymal Transition

“…The phosphorylated Smad proteins then form a trimer with Smad4 and translocate into the nucleus, where they associate and cooperate with other transcription factors to precisely regulate the target gene transcription (10). These reorchestrated target genes, as recently evidenced in several types of cancer cell lines undergoing EMT by mass spectrometry and microarray analysis, at least include the glycogenes involved in N-glycosylation, O-glycosylation, and sialylation (11)(12)(13), indicating the importance of cellular glycosylation pattern in both the transition to and the maintenance of the mesenchymal state.…”

Lectin microarray technology identifies specific lectins related to lymph node metastasis of advanced gastric cancer

“…The lectins the most significantly associated with histological type were SBA and BPL. Recently, lectin microarray analysis identified a decreased affinity for SBA and BPL, which was accompanied by hepatocyte growth factor induced epithelialmesenchymal transition of HuH7 cells, suggesting that alteration of the lectin binding profile reflecting EMT may be associated with Lauren histological type [30]. Both SMA and BPL recognize N-acetyl-D-galactosamine on the glycan structure, and this is supposed to be differentially regulated by galactosyltransferases according to histology findings in human gastric cancer [31].…”

Targeting One-Carbon Metabolism in Breast Cancer

“…Compared to resting cells, proliferating cells upregulate glycolysis and nucleotide biosynthesis pathways (Hu et al, 2013), whereas the mesenchymal metabolic signature (MMS) is enriched for glycan biosynthesis genes. Glycosylation is thought to be one of the most common covalent protein modifications in eukaryotic cells, with a major role in differentiation and mediating cell-cell interactions (Li et al, 2013). Because the EMT is accompanied by major changes in cell morphology and detachment from the surrounding cells, it is reasonable to assume that a major glycan remodeling may occur during the EMT program.…”