Hyper-O-GlcNAcylation promotes epithelial-mesenchymal transition in endometrial cancer cells

Abstract

Diabetic women have a 2–3 fold increased risk of developing endometrial cancer, however, the molecular aspects of this risk are not fully understood. This study investigated the alteration of cellular O-GlcNAcylation of proteins as the potential mechanistic connection between these two conditions. The endometrial cancer cell line (Ishikawa) was utilized to study the effect of dysregulation of O-GlcNAcylation on epithelial mesenchymal transition (EMT). Hyper-O-GlcNAcylation (via 1 μM Thiamet-G/ThmG or 25 mM Glucose) enhanced the expression of EMT-associated genes ( WNT5B and FOXC2 ), and protein expression of the EMT adhesion molecule, N-Cadherin. Reorganization of stress filaments (actin filaments), consistent with EMT, was also noted in ThmG-treated cells. Interestingly, Hypo-O-GlcNAcylation (via 50 μM OSMI-1) also upregulated WNT5B , inferring that any disruption to O-GlcNAc cycling impacts EMT. However, Hypo-O-GlcNAcylation reduced overall cellular proliferation/migration and the expression of pro-EMT genes ( AHNAK, TGFB2, FGFBP1, CALD1, TFPI2 ). In summary, disruption of O-GlcNAc cycling (i.e., Hyper- or Hypo-O-GlcNAcylation) promoted EMT at both the molecular and cellular levels, but only Hyper-O-GlcNAcylation provoked cellular proliferation/migration, and cytoskeletal reorganization.

Evidence and manipulation of O-GlcNAcylation in granulosa cells of bovine antral follicles†

“…Conversely, GCs of large follicles are presumably nearing the stages of terminal differentiation or luteinization, wherein cell division is less frequent [ 29 , 30 ], acquisition of LH receptors occurs [ 31–33 ], and resistance to apoptosis becomes attainable [ 34 ]. Indeed, under culture conditions, we and others have found that GCs typically obtained from small antral follicles exhibit greater rates of proliferation, with a shorter generation interval, and are less prone to spontaneous luteinization than GCs obtained from larger follicles [ 20 , 35 , 36 , unpublished observations and supplementary information]. In the current study, GC proliferation was measured by both mitochondrial activity (MTS assay) and the cell proliferation marker, Ki-67 (immunodetection), which is only expressed by dividing cells.…”

“…Enhancement of O-GlcNAcylation, or hyper-O-GlcNAcylation, is associated with increased cell proliferation and amplified tumorigenic potential [ 36 , 37 ]. Specifically, hyper-O-GlcNAcylation enhances cancer cell aggressiveness through increased expression of epithelial-mesenchymal transition genes, stimulating cell proliferation, and enhancing cell migration and/or invasiveness [ 36 ]. In contrast, a diminishment of O-GlcNAcylation, or hypo-O-GlcNAcylation, impairs these measures [ 36 ].…”

“…Specifically, hyper-O-GlcNAcylation enhances cancer cell aggressiveness through increased expression of epithelial-mesenchymal transition genes, stimulating cell proliferation, and enhancing cell migration and/or invasiveness [ 36 ]. In contrast, a diminishment of O-GlcNAcylation, or hypo-O-GlcNAcylation, impairs these measures [ 36 ]. Additional evidence supporting the concept that some degree of O-GlcNAcylation is necessary for cell homeostasis comes from genetic knockout experiments, in which knockout of the OGT enzyme results in an embryonic lethal phenotype [ 38 ].…”

Discovery of Spatial Peptide Signatures for Neuroblastoma Risk Assessment by MALDI Mass Spectrometry Imaging

“…AHNAK has not been previously associated with neuroblastoma, but has been implicated in several cellular functions associated with cancer, including being listed one of six putative cancer genes involved in the evolution of nine cancer types across 3000 cancer genomes [59]. Most interestingly, AHNAK has been reported to be associated with enhanced proliferation and migration in rhabdomyosarcoma [60] among other cancers as well as supporting EMT in hepatoblastoma [61], endometrial [62] and lung [63] cancer cells as well as pancreatic ductal adenocarcinoma [42] and gastric cancer [64]. A similar role in neuroblastoma would be well in line with our previous observations that several signaling elements involved in EMT regulation are mutated in relapsed neuroblastomas [13].…”

Functional Analysis of O-GlcNAcylation in Cancer Metastasis

“…For example, OGT is required for the induction and maintenance of EMT in NSCLC (84). In addition, hyper-O-GlcNAcylation contributes to the EMT of EC (85). The cell surface protein E-cadherin mediates cell-cell interactions, which is directly correlated with cancer cell adhesiveness, and this therefore mediates the invasive and metastatic capabilities of cells (86).…”