C2GnT-M is downregulated in colorectal cancer and its re-expression causes growth inhibition of colon cancer cells

Abstract

Changes in carbohydrates on the cell surface are associated with tumor malignancy. The mucin-type core 2 b-1,6-N-acetylglucosaminyltransferase (C2GnT-M) is highly expressed in the gastrointestinal tract and catalyses the formation of core 2, core 4, and blood group I branches on O-glycans. In the present study, we evaluated the role of C2GnT-M in colorectal cancer. C2GnT-M downexpression was observed in 73.6% of the primary tumors from colorectal cancer patients (39 of 53) analysed by cancer profiling array. Consistently, the majority of colon cancer cell lines and primary colon tumors expressed lower levels of C2GnT-M than did normal colon tissues by RT-PCR. HCT116 cells stably transfected with C2GnT-M inhibited expression of the core 1 structure, Galb1,3GalNAca1-Ser/Thr, on the cell surface. Moreover, C2GnT-M expression suppressed cell adhesion, motility, and invasion as well as colony formation ability. The growth of C2GnT-M-transfected HCT116 and SW480 cells was dramatically suppressed, and the cell death induced by C2GnT-M was demonstrated by an increase in the annexin V-positive cells. Interestingly, C2GnT-M inhibited cell adhesion to collagen IV and fibronectin, and decreased tyrosine phosphorylation of paxillin, indicating that the changes in cancer behavior may be partly mediated by integrin-signaling pathways. Tumor growth in vivo was also significantly suppressed by C2GnT-M in the xenografts of nude mice. These results demonstrate that C2GnT-M is frequently downregulated in colorectal cancer and suppresses colon cancer cell growth.

An integrated computational and experimental study uncovers FUT 9 as a metabolic driver of colorectal cancer

“…Interestingly, we found downregulation of the glucosidase II alpha subunit GANAB (fold change = −11.28) and Core2/Core4 beta‐1,6‐N‐acetylglucosaminyltransferase GCNT3 (fold change = −3.55) (Appendix Fig S3A). Downregulation of both these genes has been previously shown to lead to enhanced cell migration and invasion, resulting in aggressive cancers, consistent with our results showing FUT9 knockdown leads to increased colony formation and cell migration (Fig 3D and F) (Huang et al , 2006; Chiu et al , 2011). Previous studies in colorectal cancer have shown that higher levels of core 1 glycans, T antigen, and Tn antigen are the most predominantly observed O‐glycosylation changes (Holst et al , 2015).…”

Core3 O-Glycan Synthase Suppresses Tumor Formation and Metastasis of Prostate Carcinoma PC3 and LNCaP Cells through Down-regulation of α2β1 Integrin Complex

“…The decrease in ␣2␤1 integrin complex formation led to decreased FAK phosphorylation, resulting in reduced lamellipodia formation. Conversely forced expression of C2GnT-M resulted in decreases in paxillin but not FAK phosphorylation (21). Because paxillin phosphorylation likely occurs downstream of FAK phosphorylation in integrin signaling (38), C2GnT-M may affect other molecules in addition to integrin, thereby modulating paxillin phosphorylation.…”

“…1). Huang et al (21) reported that C2GnT-M is down-regulated in colonic carcinoma cells and that forced expression of C2GnT-M in HCT116 colonic carcinoma cells significantly decreased cell invasion and subcutaneous tumor formation. How up-regulation of core3 and core4 O-glycans influences the pathophysiology of cells expressing core3 and core4 O-glycans has not been addressed.…”

β1,4-N-Acetylgalactosaminyltransferase III Enhances Malignant Phenotypes of Colon Cancer Cells

“…Furthermore, overexpression of ST6GalNAc I causes a significant change in the O-glycosylation of integrin h 1 chain and leads to alterations of cell morphology and behavior (11). Recently, we found that C2GnT-M overexpression decreases colon cancer cell growth and tyrosine phosphorylation of paxillin, which is an important downstream signaling molecule of integrins (12). In addition, h3Gn-T6 was found to decrease core 1 structure and suppress metastatic potential of colon carcinoma cells through an unclear mechanism (13).…”

“…Cell adhesion assays were done according to published protocols with slight modifications (12). Ninety-six-well plates were coated with bovine serum albumin (BSA) control, human collagen IV (Sigma), human fibronectin (Sigma), or murine laminin (Sigma) at concentrations of 1 or 5 Ag/mL in PBS at 37jC for 4 h, and then blocked with 1% bovine serum albumin at 37jC for 2 h. Cells were trypsinized, washed with DMEM, and recovered in DMEM at 37jC for 40 min.…”