Effect Of S100A4 On Invasion And EMT Of CNE2 Cells

Objectives The over-expression vector of S100A4 gene was constructed to establish a stable cell line with over-expression of S100A4, to investigate the effects that the S100A4 gene was transfected into CNE2 cells. The expression levels of E-cadherin and Fibronectin which are the EMT markers and the matrix metalloproteinase MMP2 and MMP9 were detected and observed, to investigate the relationship between the expression of S100A4 gene and the invasive ability and EMT of CNE2 cells.Methods Growing CNE2 cells, obtaining cDNA and using PCR method to construct the recombinant plasmid pCMV-S100A4 which is over-express S100A4 gene. The pCMV-S100A4 was transfected into CNE2 cells by the LiposomineTM 2000. Via G418 screening, selecting the stable mono cloned transfected gene. The expression level of S100A4 gene before and after transfection was detected by Western blotting; The cells growth curve was drown by MTT method. The expression of E-cadherin and Fibronectin, MMP2 and MMP9 were detected by western blotting. Transwell chamber were used to evaluate the effects of S100A4 on the invasive ability of CNE-2 cells.Results 1. the pCMV-S100A4 was successfully constructed; 2. S100A4 was stably expressed in pCMV-S100A4-CNE2 cells; 3. pCMV-S100A4-CNE2 cells growth faster; 4. the invasion of pCMV-S100A4-CNE2 cells count increased; 5. pCMV-S100A4-CNE2 cells were low expression of E-cadherin、over expression of Fibronectin、MMP2 and MMP9.Conclusions1. S100A4 can bloost CNE2 proliferation;2. S100A4 can Participate in the EMT process in CNE2 cells.