Figure 3From: Wnt3a expression is associated with epithelial-mesenchymal transition and promotes colon cancer progressionWnt3a overexpression induced mesenchymal phenotype and increased expression of mesenchymal markers. (A) Wnt3a protein levels were significantly increased in clone7 and clone15 HCT116 cell pools transfected with Wnt3a plasmid. Then, c-myc and CyclinD1 protein expression increased in clone7 and clone15 HCT116 cell. (B) Immunofluorescent staining of F-actin. Wnt3a-overexpressing cells exhibited dramatic changes in cell morphology from a tight packed, polarized, and epithelial-like appearance to a scattered, irregular, and fibroblastic-like shape. (C) Wnt3a-overexpressing cells showed lower E-cadherin expression but higher vimentin expression. β-Catenin expression did not markedly change in three groups of cells. EMT regulatory proteins including Snail, Slug, and Twist were detected. Snail was upregulated in Wnt3a-overexpressing cells compared with control cells, whereas Slug and Twist expression did not significantly change. (D) Immunofluorescent staining of E-cadherin, vimentin, and β-catenin. Wnt3a-overexpressing cells showed lower E-cadherin expression and higher vimentin expression. More β-catenin accumulation in nucleus was observed in Wnt3a-overexpressing cells than in control cells. A green or red signal represents staining for corresponding protein, whereas a blue signal represents nuclear DNA staining by 4′,6-diamidino-2-phenylindole.Back to article page