Involvement of O-glycosylation defining oncofetal fibronectin in epithelial-mesenchymal transition process

• DOI: 10.1073/pnas.1115191108
• Publication Date: 2011-10-18T05:23:13Z
• Authors: Leonardo Freire-d...; Kirill Gelfenbeyn; Yao Ding; Ulla Mandel; Henrik Clausen; Kazuko Handa; Sen-itiroh Hakomori
• Keywords: 0301 basic medicine; Epithelial-Mesenchymal Transition; Glycosylation; Messenger; Blotting, Western; Small Interfering; Antibodies; Cell Line; Mice; 03 medical and health sciences; Transforming Growth Factor beta; Monoclonal; Animals; Humans; RNA, Messenger; RNA, Small Interfering; DNA Primers; Neoplastic; Blotting; Reverse Transcriptase Polymerase Chain Reaction; Antibodies, Monoclonal; Epithelial Cells; Cadherins; Fibronectins; Gene Expression Regulation, Neoplastic; Gene Expression Regulation; Gene Knockdown Techniques; Polypeptide N-acetylgalactosaminyltransferase; RNA; N-Acetylgalactosaminyltransferases; Western

The process termed “epithelial–mesenchymal transition” (EMT) was originally discovered in ontogenic development, and has been shown to be one of the key steps tumor cell progression metastasis. Recently, we showed that expression some glycosphingolipids (GSLs) is down-regulated during EMT human mouse lines. Here, demonstrate involvement GalNAc-type (or mucin-type) O-glycosylation process, induced with transforming growth factor β (TGF-β) prostate epithelial We found that: ( i ) TGF-β treatment caused up-regulation oncofetal fibronectin (onfFN), which defined by mAb FDC6, expressed cancer or fetal cells/tissues, but not normal adult cells/tissues. reactivity FDC6 requires addition an O-glycan at a specific threonine, inside type III homology connective segment (IIICS) domain FN. ii This change associated typical characteristics; i.e., from fibroblastic morphology, enhanced motility, decreased marker, E-cadherin, mesenchymal markers. iii up-regulated mRNA level FN containing IIICS GalNAc-T activity for peptide substrate onfFN epitope. iv Knockdown GalNAc-T6 T3 inhibited TGF-β–induced process. v Involvement GSLs detectable these These findings indicate important functional role onfFN, site-specific domain,