Effect Of Baicalein On Proliferation,Apoptosis And Epithelial-Mesenchymal Transition Of Human Synovial Sarcoma SW982 Cells And Its Mechanism

Objective:To investigate the effect of Baicalein(BAI)on the proliferation,apoptosis and migration of human synovial sarcoma SW982 cells and its related molecular mechanism.Methods:SW982 cells were treated with BAI at concentrations of 0μM,15μM,25μM,50μM,75μM,and 100μM for 24 h,48h,72 h,and 96 h,the cell proliferation ability was detected by MTT assay and the medication concentration of BAI was screened;SW982 cells were treated with BAI for two weeks,and the colony-forming ability of the cells was detected by plate colony formation experiment;SW982 cells were treated with BAI for 48 h,and the apoptosis of SW982 cells was analyzed by Annexin V-FITC/PI double staining by flow cytometry;SW982 cells were treated with BAI for 0h,24 h,48h and 72 h,and lateral migration was detected by scratch healing assay;SW982cells were treated with BAI for 48 h,and the expression of apoptosis-related proteins(Bcl-2,Bax,Caspase-3,Cleaved-Caspase 3),EMT-related proteins(E-Cadherin,Vimentin,Snail),MMP family proteins(MMP2,MMP9)and PI3K/Akt/mTOR signaling pathway-related proteins were successfully detected by western blot.Results:1.MTT assay results showed that: SW982 cells were treated with different concentrations of BAI for 24 h,48h,72 h and 96 h,compared with the control group,the proliferation ability of SW982 cells in the BAI medication group was significantly inhibited in a concentration-and time-dependent manner(P<0.01),and BAI of 0μM,15μM,25μM and 50μM was selected for follow-up experiments.2.The results of plate colony formation assay showed that: After treatment with different concentrations of BAI,the colony forming ability of SW982 cells in the BAI medication group was significantly inhibited in a concentration-dependent manner as compared with the control group(P<0.01).3.The results of Annexin V-FITC/PI double staining by flow cytometry showed that: After the SW982 cells were treated with BAI for 48 h,the apoptosis rate of SW982 cells of 0μM BAI(control group)was 15.9%,while the apoptosis rate was as high as 31.6% when the concentration of BAI was 50μM.Compared with the control group,the apoptosis rate of SW982 cells in the BAI medication group increased in a concentration-dependent manner(P<0.01).4.The scratch healing experiment showed that: Compared with the control group,the horizontal migration ability of SW982 cells in the BAI medication group was significantly inhibited at 48 h and 72 h in a concentration-dependent manner(P<0.01).5.The western blot results showed that:(1)Compared with the control group,the expression levels of pro-apoptotic proteins Cleaved-caspase 3 and Caspase-3 of SW982 cells in the BAI medication group were significantly increased,while the expression levels of anti-apoptotic protein Bcl-2 and Bcl-2/Bax ratio were significantly decreased(P<0.05 or P<0.01);(2)Compared with the control group,the expression levels of Vimentin,Snail,MMP9 and MMP2 proteins of SW982 cells in the BAI medication group were significantly decreased,while the expression level of EMT marker protein E-Cadherin was significantly increased(P<0.05 or P<0.01);(3)Compared with the control group,the expression levels of the PI3K/Akt/mTOR signaling-related proteins p-PI3 K,p-Akt,and p-mTOR of SW982 cells were significantly down-regulated in the BAI medication group(P<0.05 or P<0.01),but the expression levels of related total proteins had no significantly change.Conclusion:1.BAI inhibited the proliferation,colony formation,migration and EMT process of SW982 cells and induced apoptosis;2.The effects of BAI on proliferation,apoptosis and migration of SW982 cells were related to its regulation of PI3K/Akt/mTOR signaling pathway.