The results together suggested that elevation of O-GlcNAcylation could enhance migration/invasion ability of CCA cells by sequential reactions as defined in this study: 1) increasing of cellular O-GlcNAcylation increases the O-GlcNAcylation of transcriptional factor NF-κB and the signaling protein (Akt); 2) O-GlcNAc modification activates the nuclear translocation of NF-κB and the phosphorylation of Akt; 3) the transcriptional activation of downstream signaling increases MMP expression and results in 4) the increasing of cell migration and invasion. The gene discussed is AKT1; the disease is cholangiocarcinoma.