ABSTRACT
Objective To investigate the role of NRAGE subcellular localization in the EMT and radioresistance of esophageal cancer cells.Methods EMT model cells were established by the treatment of TE13 cells with TGF-β1.To verify the establishment of EMT model and the phenotype of EMT-like TE13R120 cells,EMT marker mRNA and protein were detected by Real-time PCR and Western blot,respectively.Real-time PCR was also used to detect the expression of NRAGE mRNA in three groups.Total NRAGE protein,cytoplasm protein and nuclear protein were measured by Western blot.Results It was found that TGF-β1 could induce morphological alterations of TE13 cells from epithelial to mesenchymal and change the expressions of EMT maker E-cadherin and vimentin (t =13.56,-232.84,P ＜ 0.05),indicating the successful establishment of EMT model cells.Similar expression trends of EMT makers were observed in TE13R120cells (t=15.84,-54.54,P＜0.05).NRAGE mRNA (t=-8.73,-5.62,P＜ 0.05) and total protein in both EMT model cells and TE13R120 cells were higher than that in TE13 cells,especially for the nuclear proteins.However,no differences in NRAGE cytoplasm protein expression were found among the three groups.In addition,there were also no difference of NRAGE mRNA (t =-0.88,P ＞0.05),cytoplasm and nuclear protein between TE13R120 cells and EMT model cells.Conclusions The radioresistant cell line TE13R120 has the EMT-like phenotype that may cause cell radioresistance by changing the subcelluar localization of NRAGE.