Hederagenin Disrupts TGF-β1 Mediated Epithelial-mesenchymal Transition Of Grastric Cancer Cells Via The PI3K/AKT Pathway

Objective:To investigate the effect of hederagenin as a tumor suppressor on cell apoptosis,viability,migration,invasion,and cell cycle arrest.To determine if HG may be utilized for the treatment of GC,we elucidated its molecular mechanism underlying its inhibition of TGF-β1-induced EMT in relation to the PI3K/AKT pathway.Method:(1)Different concentrations of hederagenin(0,12.5,25,50,100,and 200 μg/mL)were used to intervene gastric cancer cells for 12 h,24 h,and 48 h.The cell proliferation was measured by MTT assay.(2)Evaluated the effects of HG on GC cells by using Annexin V-FITC and observed the nuclear morphological changes of cells by using Hoechst 33258 staining;apoptosis-related genes(Bcl-2,Bax,PARP,cleaved-PARP)were detected by western blot analysis.(3)The cell cycle was measured by flow cytometry.(3)Transwell assay were used to observe the changes of Migration and invasion ability of GC cells.(4)TGF-β1 was used to induce EMT in gastric cancer cells,then,HG were used to intervene.(5)Western blot was used to analyze the expression of epithelial phenotype marker protein E-cadherin,N-cadherin,Vimentin and MMP-2,MMP-9.(6)In order to verify the HG through the PI3K/AKT signaling pathway to play a regulatory role in TGF-β1-induced EMT.We used PI3K/AKT inhibitor LY294002 to confirm the effects of HG.Result:HG inhibit the proliferation,migration and invasion,promote cells apoptosis,as well as triggers S-phase cell cycle arrest.Hederagenin can attenuates TGF-β1-induced EMT by increased the expression of E-cadherin;decreased the expression of N-cadherin,Vimentin.MMP-2 and MMP-9.HG inhibits the TGF-β1-induced EMT process in GC cells via the PI3K/AKT signaling pathways.Conclusions:Hederagenin influences the biological activities of gastric cancer cells and disrupts TGF-β1-mediated epithelial-mesenchymal transition via the PI3K/AKT pathway.Conclusion:Our discovery demonstrates that hederagenin disrupts many biological activities of GC cells.Our major findings concerning Hederagenin are as follows:(1)HG inhibits proliferation,induces cell apoptosis,and arrests cells at the S-phase;(2)HG disrupts migration and invasion of GC cells without compromising cell viability;(3)HG inhibits the TGF-β1-induced EMT process in GC cells via the PI3K/AKT signaling pathways.