Effect Of Knocking Down For Smurf1 Gene On Biological Behavior Of Colon Cancer Cell Lines

Objectives:1、To construct a eukaryotic expression vector of Smurfl shRNA.2、To analyze the effect of proliferation, migration, invasion and EMT by knocking down for smurfl gene in colon cancer cell lines.Methods:We synthesized a couple of coding smurfl single-stranded shRNA sequences in vitro by reference reported. The double-stranded by annealing was inserted into pSilence 4.1-CMV vector digested by BamH I and HindⅢ. The recombination plasmid was transfected into HCT116 and SW480 by lipofection. After screening by G418, the effect of proliferation, migration, invasion and EMT was detected by MTT assay, wound healing, transwell and western blot in colon cancer cell lines.Results:1、The recombinant plasmid was constructed completely and applied to transfection.2、The cell lines that contained the recombination plasmid were at low proliferation rate in compare with control group.3、The ability of migration was decreased in the cell lines with the recombination plasmid.4、After transfection, the interference group was reduced in cell number through membrane.5、Down-regulated expression of smurfl gene increased the level expression of E-cadherin, meanwhile decreased the expression level of Vimentin and Twist.Conclusions:1.We constructed successfully the plasmid and the expression level of smurfl was reduced by it.2.After expression level of smurfl decreased, the ability of proliferation, migration and invasion was reduced, and it reversed the mesenchymal-like morphological changes, inhibited epithelial-mesenchymal transition.