Metformin Synergizes With Thioridazine To Enhance The Inhibitory Effect And Mechanism Of Lung Cancer

Objective: Explore the effects of metformin and thioridazine on the proliferation,migration and invasion of non-small cell lung cancer,determine whether the two drugs have a synergistic effect,and further explore their mechanism of action.Methods: Lung cancer cells were cultured in vitro,and human lung adenocarcinoma cell lines H1975 and human lung squamous cell carcinoma cell lines H520 were used as the research objects to observe the effects of thioridazine,metformin,and the combination of the two drugs on the proliferation,plate clone formation,migration and invasion of H1975 and H520 cells.The MTT method was used to detect the effects of different concentrations of thioridazine and metformin on the proliferation of H1975 and H520 cells at 24 h and48h;Detect the effects of thioridazine and metformin alone and the combination of the two drugs on the cloning ability of H1975 and H520 cells through the plate clone formation experiment;Transwell migration and invasion experiments are used to detect the effects of the two drugs alone and in combination on the migration and invasion of H1975 and H520cells;Western blot was used to detect the effect of thioridazine,metformin and the combination of the two drugs on the expression of PI3K/Akt/mTOR pathway-related proteins;Detect the changes of p53,Bax,Bcl-2,Caspase-3,Caspase-9 protein expression levels after the action of thioridazine and metformin.Results: Through the MTT experiment,it was found that thioridazine and metformin inhibited the proliferation of lung cancer cells H1975 and H520 in a concentration-dependent manner.And the two drugs have a synergistic(sensitization)effect.Compared with the control group,thioridazine,metformin and the combination of the two drugs can inhibit the clonal formation,migration and invasion of H1975 and H520 cells.However,compared with the single-agent group,the combined drug group had a significantly enhanced tumor suppressor effect.Combined use of thioridazine and metformin inhibits the proliferation,migration,invasion and apoptosis of lung cancer cells.It is related to down-regulating the expression of p-Akt,p-PI3 K,p-mTOR,and Bcl-2protein,and up-regulating the expression level of p53,Bax,Caspase-3,and Caspase-9.Conclusion: The inhibitory effect of metformin synergistically sensitizing thioridazine on lung cancer cells is related to down-regulation of p-Akt,p-PI3 K,p-mTOR,Bcl-2 and up-regulation of p53,Bax,Caspase-3,and Caspase-9 protein expression.