Specific N-glycan alterations are coupled in EMT induced by different density cultivation of MCF 10A epithelial cells.

Abstract

Epithelial-mesenchymal transition (EMT) is a process in tumor progression during which cancer cells undergo dramatic changes acquiring highly invasive properties. In a widespread adoption of TGF-β-induced EMT model, we have previously observed that expression of bisecting GlcNAc on N-glycans was dramatically decreased. Herein, we performed in vitro studies with the MCF10A cell line. In response to low cell density, MCF10A cells suffered spontaneously morphologic and phenotypic EMT-like changes, including elongated spindle shape, extended out from edge of the cell sheet, cytoskeleton reorganization, vimentin and fibronectin up-regulation, catenins redistribution, and cadherin switching. Moreover, these phenotypic changes were associated with specific N-glycan alterations. Interestingly, the amounts of bisecting GlcNAc structure were declined, by contrast, the formation of β1-6 GlcNAc branches were obviously up-regulated during the EMT induced by sparse cell conditions. We further investigated N-glycans on the β1-integrin, which is a good target of some glycosyltransferases. The reactivity with E4-PHA lectin decreased, whereas the staining for L4-PHA lectin, which recognizes branched GlcNAc, increased in sparse cell conditions compared with dense cell conditions. Taken together, these results demonstrated that specific N-glycan alterations are coupled in EMT process and promoted cells migration at a low cell density.