Overexpression Of TGM1 Inhibits Epithelial-Mesenchymal Transition In Breast Cancer Cell FE1.2

Objective: To explore the effects of the TGM1 gene on Epithelial-Mesenchymal Transition(EMT)in breast cancer cells.Methods: Q-RT-PCR was used to detect the expression levels of TGM1 gene in rat FE1.2 and FE1.3 breast cancer cells.The expression vector p CMV3-TGM1 was then transfected into FE1.2 cells.Subsequently,the morphological changes of FE1.2 cells after transfection with p CMV3-TGM1-GFPspark was observed under microscope.The effect of TGM1 overexpression on the proliferation of FE1.2 cells was examined by MTT.FE1.2-TGM1 and FE1.2-vector cells were then tested for changes in m RNA and proteins expression.The expression of EMT molecular markers E-cad,Vim and Cyclin-D1,?-catenin were then examined by Western blotting and Q-RT-PCR on these cells.flowcytometry method was used to detect changes in cell particle size.Finally,the migration ability was showed by wound healing assay.Results: The expression of TGM1 in breast cancer cell line FE1.3 was significantly higher than that in FE1.2 cells.after transfection of recombinant vector p CMV3-TGM1 into FE1.2 cells,TGM1 expression was significantly upregulated(m RNA: P <0.001,protein:P <0.01).The shape of FE1.2 cells after transfection with with p CMV3-TGM1-GFPspark was changed.;As compared with the blank control and vector group,the proliferation ability of FE1.2 cells transfected with p CMV3-TGM1 was significantly inhibited(24h: P<0.01,48 h and 96h: P<0.001).The m RNA level of E-cad was up-regulated(p <0.01)and Vim levels was downregulated(P <0.001).In these cells,the m RNA level of cyclin-D1 was significantly up-regulated(P <0.01).?-catenin and Vim expression was downregulated(P <0.01).The flow cytometry analysis showed a change in cell size when compared to control.Wound healing assay shows that the migration ability was significantly inhibited(P<0.05).Conclusion: Over-expression of TGM1 can inhibit EMT in breast cancer cells.