The Effects And Mechanisms Of Plumbagin Comebined With PF Preoperative Chemotherapy On Apoptosis And Autophagy In Tongue Squamous Cell Carcinoma

Objective:To investigate the effects and mechanisms of plumbagin comebined with PFpreoperative chemotherapy on apoptosis and autophagy in tongue squamous cellcarcinoma.Methods:1.Human tongue squamous cell carcinoma CAL27 cells were cultured in vitro.2.MTT assay was applied to examine the proliferation inhibition effect and halfmaximal inhibitory concentration(IC50)of plumbagin,cisplatin and 5-fluorouracil on CAL27 cells.MTT assay was applied to examine the proliferation inhibition effect of plumba-gin with PF regimen chemotherapy on CAL27 cells.3.Flow cytometry was employed to detect the apoptosis level in CAL27 cells afterthe treatment of plumbagin and/or PF regimen chemotherapy.4.Cyto ID flow cytometry was employed to detect the autophagy level in CAL27cells after the treatment of plumbagin and/or PF regimen chemotherapy.5.Western blot was used to examine the expressing level of the autophagy relatedproteins Beclin1 and LC3 after the treatment of plumbagin and/or PF regimen chemotherapy.6.Western blot was used to examine the expressing level of the signaling pathwayproteins after the treatment of plumbagin and/or PF regimen chemotherapy.Results:1.Plumbagin,cisplatin and 5-fluorouracil can inhibit the proliferation of CAL27 cells via concentration dependent manner,the IC50 in 24 h were 7.77?mol/L,10.50?g/ml,1.26mg/ml separately.2.When CAL27 cells were treated by the combination group of PB with PF,the proliferation was significantly higher than the PF single group.3.When CAL27 cells were treated by the combination group of PB with PF,the apoptosis ratio was significantly higher than the PF single group.4.When CAL27 cells were treated by the combination group of PB with PF,the autophagy level was significantly higher than the PF single group.5.When CAL27 cells treated with PB alone,the expressing level of PI3 K,p-AKT,and p-mTOR proteins were decreased at different degrees,while the combination treatment group had the same results than the PF single group.6.After treatment with IGF-1 and MHY1485,the down-regulation effect of PI3 K,p-AKT and p-mTOR proteins by PB was reversed,at the same time,it significantly inhibited the induction of apoptosis and autophagy by PB in combination treatment group in CAL27 cells.Conclusions:Plumbagin induces apoptosis and autophagy via inhibiting PI3K/AKT/mTOR signaling pathway to enhance the killing of PF regimen chemotherapy in tongue squamous cell carcinoma CAL27 cells.