The Study On The Role And Mechanism Of FK866 In EMT Of Hepatocarcinoma MHCC97-H Cells In Regulating NAMPT/NAD~+/SIRT1 Signal Pathway

Objective:HCC cell invasion and migration are two of the most clinically important characteristics of the tumor,and EMT serves a crucial function in these processes.Therefore,finding new targeting drugs of EMT has become the current research focus.Current studies demonstrated that FK866,a targeted NAMPT inhibitor,inhibits the viability of HCC cells and induces cancer cell apoptosis;however,the effect of FK866 on the invasion and metastasis of HCC cells remains unknown.In this study,small molecule targeting drug FK866 was used to treat hepatocellular carcinoma MHCC97-H cells to observe the changes of cell function,and to explore the effects of FK866 on the proliferation,invasion and metastasis of human hepatocellular carcinoma MHCC97-H cells and the possible mechanisms.Method:The human hepatocellular carcinoma MHCC97-H cells were treated with different concentrations of FK866 in vitro and divided into control group and experimental group;CCK8 experiment was used to observe the changes of cell viability after FK866 treated cells for 24h,48h and 72h;The levels of NAD~+and ATP in MHCC97-H cells were measured using the Microdetermination assay kit;The effects of FK866 on the invasion and metastasis of MHCC97-H cells were investigated using wound healing,invasion and migration assays;Western blot analysis was performed to detect SIRT1,Vimentin and E-Cadherin expression in FK866 treated MHCC97-H cells.Results:The results of CCK8 experiment suggest that FK866 inhibits the viability of MHCC97-H cells in a dose-dependent(P<0.05);The present study indicates that FK866decreases the levels of NAD~+and ATP in MHCC97-H cells in a dose-dependent(P<0.05);We find that FK866 inhibits the invasion and migration of MHCC97-H cells by using wound healing,invasion and migration assays in a dose-dependent(P<0.05);The results of the present study indicated that the expression of SIRT1 and Vimentin was downregulated significantly and that of E-cadherin was upregulated significantly at the protein level(P<0.05).Conclusion:1.FK866 can inhibit the viability of the HCC cell line MHCC97-H;FK866can decrease the levels of NAD~+and ATP in MHCC97-H cells.2.FK866 can inhibit the invasion and migration of MHCC97-H cells.3.By inhibiting the NAMPT/NAD~+/SIRT1 pathway,FK866 can up-regulate E-cadherin and down-regulate Vimentin to reverse the EMT process of HCC and further inhibit the invasion and metastasis of HCC.