Effect And Mechanism Of Pirfenidone On Epithelial-Mesenchymal Transition Induced By Cancer-Associated Fibroblasts In HT29 Colon Cancer Cell Lines

Aim: To investigate the effect and mechanism of pirfenidone on the invasion,migration,proliferation and epithelial-mesenchymal transition(EMT)induced by cancer-associated fibroblasts(CAFs)in HT29 colon cancer cell lines.Methods: CCK-8 was used to measure HT29 proliferation efficiency after co-culture of CAFs with HT29 using co-culture chamber.The transcription and protein expression levels of E-cadherin and Vimentin in HT29 colon cancer cells were detected by Real time-PCR and Western blot,respectively.Trans-well assay was used to detect the migration and invasion ability of HT29.ELISA technique was used to detect the expression of cytokines in CAFs supernatant.Results: 1.The concentration of PFD of 0.25 and 0.5mg/ml had no inhibitory effect on the proliferation of HT29 cells.Pirfenidone inhibited the proliferation of HT29 induced by CAFs(P <0.05).2.The expression of Vimentin was significantly increased in HT29 cells cultured in CAFs conditioned medium(P <0.05),while the expression of E-cadherin was significantly decreased(P <0.05).However,Conditioned media of pirfenidone treated CAFs had no significant effect on the expression of the two proteins in HT29 cells(P >0.05).3.Trans-well assay showed that after co-cultured with CAFs,the invasion and migration ability of HT29 were significantly enhanced(P <0.05).4.The results of ELISA showed that the secretion of TGF-β1 by CAFs was inhibited by pirfenidone,which was dose related.Conclusion: CAFs can induce HT29 colon cancer cell lines epithelial-mesenchymal transition and enhance their invasion/migration ability.Pirfenidone inhibits the invasion,migration,proliferation and epithelial-mesenchymal transition of HT29 colon cancer cell lines by inhibiting the secretion of TGF-β1 by CAFs.