Effects Of Silencing Expression Of FoxM1 Gene On Epithelial-mesenchymal Transition In ECA-109 Esophageal Squamous Cell Carcinoma

Objective:To observe the effects of silencing of transcription factor FoxM1 in Epithelial-mesenchymal transition(EMT) of ECA-109 Esophageal Squamous Cell Carcinoma,FoxM1 gene was reduced to explore its influence on migration of ECA-109 Esophageal Squamous Cell Carcinoma and to explore preliminarily mechanism of it in EMT.Methods:FoxM1 plasmid was transfected into ECA-109 cells for 48 hours.The expression of FoxM1 gene and epithelial mesenchymal markers E-cadherin and Vimentin were detected by reverse transcription-polymerase chain reaction(RT-PCR) and Western blot after transfection 48 hours,the strongest interference group was selected to carry out the formal experiment.FoxM1-shRNA and NC-shRNA vector was transfected into ECA-109 cells by lipofectamine2000,to set up three groups of working cell line as follows:No transfection of ECA-109 cells?NC-shRNA?FoxM1-shRNA.The expression of FoxM1? E-cadherin and Vimentin gene were detected by RT-PCR and Western blotting in three group cells.we detected the effect of down-regulation of FoxM1 gene on migration in ECA-109 cells.Results: 1.Filtering the interference efficiency of FoxM1-shRNA in ECA-109 cells.Compared to control group and negative control group NC-sh RNA,Western-blotand RT-PCR results showed that the interference efficiency of shRNA2 was obviously higher than others(p<0.01). So we chose interference carrier shRNA2 for formal experiment.2.Influence on the expression of epithelial mesenchymal markers E-cadherin and Vimentin in ECA-109 cell transfected FoxM1-shRNA2.Compared to control group and negative control group NC-sh RNA,the expression of FoxM1 in FoxM1-shRNA2 transfection group was decreased(p<0.05),the expression of Vimentin in FoxM1-shRNA2 transfection group was decreased(p<0.05), while the expression of E-cadherin in FoxM1-shRNA2 transfection group was increased(p<0.05) by Western-blot.3.The influence of down-regulation of FoxM1 gene on migration in ECA-109 cellsScratch results showed that control group and negative control group NC-shRNA,the cells were exposed to regional migration after 24 hours,then the cells of control group and negative control group were covered in the middle,While the scratch was still visible in FoxM1-shRNA2 transfection group.Conclusion: 1.The down-regulation of FoxM1 gene could inhibit the ECA-109 cells converting from normal epithelial phenotype cell to mesenchymal phenotype cell.2.The down-regulation of FoxM1 gene reduced the migration capabilities in ECA-109 cells.