Effects Of Capsaicin On Proliferation、Migration And Autophagy Of Choroidal Melanoma Cell Lines C918 And OCM-1

Purpose:To search the effects of capsaicin on the proliferation,migration and autophagy of choroidal melanoma cell lines C918 and OCM-1,and to initially investigate the underlying mechanisms.Methods:The effects of three concentrations 100μmol/L,200μmol/L and 300μmol/L of capsaicin on the proliferation of C918 and OCM-1 cells were tested by CCK8 and plate cloning experiments.The effects of three concentrations of capsaicin on the migration ability of C918 and OCM-1 cells were tested by cell scratch test,Transwell cell migration test,and the expression of mesenchymal marker proteins Vimentin and N-cadherin.The effect of three concentrations of capsaicin on autophagy of C918 and OCM-1 cells was detected by the expression of autophagy marker protein LC3-Ⅱ.The effects of three concentrations of capsaicin on the AMPK/mTOR signaling pathway in C918 and OCM-1 cells were detected by the expression of p-AMPK and p-mTOR proteins.Immunofluorescence experiments were performed to detect LC3-Ⅱprotein expression in C918 and OCM-1 cells.Complete medium with 100μmol/L,200μmol/L and 300μmol/L capsaicin was used as the experimental group.Complete medium without capsaicin was used as the control group.All experiments were repeated three times,and the statistical method was t-test.Results:1)CCK-8 results showed that three concentrations of capsaicin inhibited cell viability of c918 cells(P<0.0001);However,when capsaicin was applied to OCM-1cells,only at a high concentration of 300 μmol/L capsaicin caused a significant decrease in cell viability(P<0.01).Plate cloning experiments showed that 100,200,300 concentrations of capsaicin can reduce the number of cell clones in C918 cells(P<0.05;P<0.001;P<0.0001).Plate cloning experiments showed that 100,200,300 concentrations of capsaicin can reduce the number of cell clones in OCM-1 cells(P<0.001;P<0.0001;P<0.0001).2)In the scratch test,the cell mobility at 200μmol/L and 300μmol/L capsaicin group for 24 h decreased(P<0.001;P<0.0001).The cell mobility at 200μmol/L and300μmol/L capsaicin group for 48 h decreased(P<0.05;P<0.0001).The cell mobility at 300μmol/L capsaicin group for 96 h decreased(P<0.0001).The results of Transwell cell migration assay showed that three concentrations of capsaicin could reduce the number of membranes penetrated by C918 cells and OCM-1 cells(P<0.0001).The results of Western blot experiments showed that capsaicin at a concentration of300μmol/L reduced vimentin expression in c918 cells(P<0.01);Capsaicin at200μmol/L and 300μmol/L concentration reduced N-cadherin expression in C918cells(P<0.05;P<0.01).Western blot showed that capsaicin at 200μmol/L and300μmol/L concentrations can reduce the expression of Vimentin in OCM-1 cells(P<0.05;P<0.0001);Capsaicin at a concentration of 300μmol/L can reduce the expression of N-cadherin in OCM-1 cells(P<0.05).3)Western blot results showed that the capsaicin concentrations at 200μmol/L and 300μmol/L increased the expression of LC3-Ⅱ protein in C918 cells(P<0.05;P<0.05);Capsaicin at 100μmol/L,200μmol/L and 300μmol/L concentrations of capsaicin could increased the expression of LC3-Ⅱ protein in OCM-1 cells(P<0.05;P<0.01;P<0.01).4)Western blot also showed that capsaicin at a concentration of 300μmol/L can increased the expression of p-AMPK in C918 cells(P<0.001);The capsaicin concentrations at 200μmol/L and 300μmol/L decreased the expression of the expression of the p-mTOR in C918 cells(P<0.05;P<0.05).The capsaicin concentrations at 100μmol/L,200μmol/L and 300μmol/L increased the expression of p-AMPK in OCM-1 cells(P<0.05;P<0.01;P<0.01).Three capsaicin concentrations decreased the expression of the p-mTOR in OCM-1 cells(P<0.0001).Conclusion:Capsaicin inhibited the proliferation of C918 and OCM-1 cells.Capsaicin inhibited the migration ability of C918 and OCM-1 cells.Capsaicin could promote autophagy in C918 and OCM-1 cells.Capsaicin may participate in the proliferation,migration,and autophagy of C918 and OCM-1 cells by regulating the AMPK/mTOR signaling pathway.