The Effect Of Silencing Maspin On The Epithelial-mesenchymal Transition And The Migration、Invasion Ability In T24 Cells

Objective The research choosed the bladder carcinoma T24 cells as the research object to observe the effect of silencing maspin gene by RNAi on the EMT and the migration、invasion ability.Methods The bladder carcinoma T24 cells were cultured,the interference plasmid maspin-shRNA and empty plasmid was established and transfected into T24 cells.Then the blank control was designed.After transfection for 48 h,the expression mRNA and protein of maspin were detected,and the most effective interference group were selected.The experiment were divided into 3groups as follows:T24group,T24+N.C shRNA group and T24+maspin-shRNA group.After 48 h changes in cell morphology were observed by microscope;The expressions of EMT markers E-cadherin and vimentin were detected by Western Bloting,and the effect of EMT of T24 cells after down-regulation of maspin gene were observed;The T24 cells abilities of migration and invasion after down-regulation of maspin were detected by wound scratch and transwell assays.Results The interference efficiency of shRNA-4 was obviouslyhigher than other 3groups(shRNA-3,shRNA-5,shRNA-6).Compared with negative control group,most cells of maspin-shRNA group became more loose with larger intercellular space,and the cells morphology became fusocellular,in a word,phenotype of the cells became mesenchymal.According to result of Western Bloting,the expression of Vimentin increased and the expression of E-cadherin decreased(P<0.01).Wound scratch and transwell assay showed that the migration and invasive ability of T24 cells were increased after down-regulation of maspin.(P<0.05).Conclusion Silencing maspin can promote the development of EMT in bladder carcinoma T24 cells;Silencing maspin can enhance the ability of the migration and invasion in T24 cells,it may be related with the development of EMT by silencing maspin in T24 cells.