The Research Of Wnt3a And Wnt3Protein Induced EMT In Mouse Hepatic Progenitor Cells

ObjcetiveThe recombinate adenovirus Ad-GFP-wnt were trancfected in mousehepatic progenitor cells to obtain highly expressed wnt gene. Throughobserved the change of epithelial-mesenchymal transition and migrationabilities in highly expressed wnt gene’s mouse hepatic progenitor cells,approach the effect of wnt signaling pathway on EMT in mouse hepaticprogenitor cells.MethodsAd-GFP-wnt virus were transfected in mouse hepatic progenitorcells.Observed the sharp changes with blank comparedgroup.Wound-healing assay and transwell assay were used to observed themigration abitities.Realtime-PCR and western blot were used to detect theexpression of epithelial and mesenchymal markers respectively.ResultsWnt3a induced epithelial-mesenchymal transition and enhanceimgaration ability in mouse hepatic progenitor cells 1.Microscopic observation showed that mouse hepatic progenitor cellswere suffered irregular polygons to long spindle.Would-healing assayshowed that the distance of Ad-wnt3a and Ad-GFP were （0.53±0.05）mmand（0.33±0.02）mm respectively, the migration ability were inhanced inAd-wnt3a group compared with the blank control group.Transwell assayalso found that cells number of Ad-wnt3a group and Ad-GFP group wentthrough venticles were （62.00±3.60）and （10.33±2.08）.The cells numberwere increased compared with the blank control group.2.Realtime-PCR and Western-blot showed that the expression ofmesenchymal markers N-cadherin,vimentin,snail1in mRNA and proteinlevel were increased and the expression of epithelial markers weredecreased.Wnt3promote the role of hepatic progenitor cells inEpithelial-mesenchymal transition1.Observed under microscope we can see the normal14-19cells wereepithelial characteristic sharp while high expression wnt3were spink-likesharp.Would-healing assay and transwell assay showed that the highexpression wnt3group’s migration abillties were improve significantlycompared with blank control group.2.Realtime-PCR and western-blot were found that mesenchymalmarkers were incrased in mRNA and protein level,while epithelial markerswere decrased compared with blank control group. conclusion1.Successfully constructed high expression wnt gene in liverprogenitor cell lines.2. wnt3a can induce epithelial-mesenchymal transformation andenhance their ability to migrate in mouse hepatic progenitor cells.3.wnt3can promote mouse liver progenitor cells of epithelialmesenchymal transition, and improve their migration ability4.wnt signaling regulates EMT conversion process in mouse hepaticprogenitor cells, suggesting that it may be involved in the process of liverfibrosis.