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Here, we demonstrated the involvement of <jats:italic>O\u2010<\/jats:italic>GlcNAcylation in melanoma metastasis. Using the data from GEO database, we found that <jats:italic>O\u2010<\/jats:italic>GlcNAcylation and its related enzymes, including glutamine fructose\u20106\u2010phosphate amidotransferase (GFAT), <jats:italic>O\u2010<\/jats:italic>GlcNAc transferase (OGT), and <jats:italic>O\u2010<\/jats:italic>GlcNAcase (OGA); were elevated in metastatic melanoma compared with primary tumors and normal tissues. Functional analyses in melanoma cell lines \u2013 MNT\u20101, SK\u2010MEL\u201028, and A\u2010375 showed that suppression of <jats:italic>O\u2010<\/jats:italic>GlcNAcylation by siRNA against OGT significantly reduces the migration and invasion abilities of the cells. Phosphorylation of Akt and NF\u03baB was drastically suppressed after the knockdown of OGT, suggesting the role of <jats:italic>O\u2010<\/jats:italic>GlcNAcylation in regulating the Akt\u2010NF\u03baB signaling pathway. In addition, we found that the NF\u03baB target genes, such as ZEB\u20102 and MCT\u20101, were significantly upregulated in metastatic tumors compared with primary tumors. MCT\u20101 expression in melanoma tissues was also correlated with the <jats:italic>O<\/jats:italic>\u2010GlcNAcylation level. Taken together, we have demonstrated in this study the possible role of <jats:italic>O\u2010<\/jats:italic>GlcNAcylation in controlling melanoma metastasis via upregulating MCT\u20101 expression through activation of the Akt\u2010NF\u03baB signaling pathway.<\/jats:p>","DOI":"10.1002\/cbic.202400896","type":"journal-article","created":{"date-parts":[[2025,1,29]],"date-time":"2025-01-29T07:10:43Z","timestamp":1738134643000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Silencing of <i>O\u2010<\/i>GlcNAc Transferase Attenuated <i>O\u2010<\/i>GlcNAcylation and Metastatic Potentials of Melanoma Cells Through Suppression of Akt\u2010NF\u03baB Signaling Pathway"],"prefix":"10.1002","volume":"26","author":[{"given":"Nopkamol","family":"Kanchanangkul","sequence":"first","affiliation":[{"name":"Department of Biochemistry Faculty of Medicine Khon Kaen University  Khon Kaen 40002 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