Figure 4. Regulation the SIRT1 expression in chondrosarcoma cells leads to the change of epithelial-to-mesenchymal transition level.

(A and B) SW1353 and HS.819. T cells (1 × 10⁶/well) were cultured in 6-well plate for 24 h with the existence of Ad-SIRT1 or Ad-shSIRT1. Then the cells were collected and realtime PCR was employed to examine the expression of EMT associated genes including Vimentin, N-cadherin, Twist, E-cadherin and β-catenin. (C) Immunofluorescent staining of E-cadherin and Vimentin was performed in SW1353 HS.819.T cells, nuclei were counterstained with DAPI. The pictures were captured by fluorescence microscopy (×200). (D) Western blot was performed to detected Twist expression. (E) The Twist in SW1353 cells was inhibited by shRNA and then the metastasis potential of the cells was examined by transwell assay. The cells were cultured in the upper chamber of the transwell for 48 hours in serum-free medium, while 5% fetal bovine serum was placed in the lower chamber. The number of cells which has invaded into matrigel was counted in 10 random fields under microscope (×200; *P < 0.05).