Mechanisms Of Luteolin Inhibiting Proliferation、Adhesion、Migration And Invasion Of Choroidal Melanoma Cells

Purpose:To investigate the effects of luteolin on the proliferation,apoptosis,adhesion,migration and invasion of choroidal melanoma cells C918 and OCM-1 in vitro and their related mechanisms.Methods:CCK-8 assay was used to determine the time and concentration of luteolin on C918 cells and OCM-1 cells.The ability of cell proliferation was detected by plate clone formation assay and Ed U cell proliferation assay.Hoechst staining and AnnexinⅤ-FITC/PI double staining flow cytometry were used to detect apoptosis.Matrigel cell adhesion assay,wound scratch assay,Transwell cell migration assay and Transwell cell invasion assay were used to detect the ability of cell adhesion,migration and invasion.The expression of MMP-2 was detected by ELISA.Western blot was used to detect the expression of p-PI3 K P85,Akt,and p-Akt proteins,and the expression of cytoskeleton protein Vimentin was also detected.C918 cells and OCM-1 cells were pretreated with IGF-1 and then treated with luteolin to detect the role of luteolin in inhibiting cell clone formation,adhesion,migration,invasion and the correlation of PI3K/Akt signaling pathway.Results:The CCK-8 assay results showed that the IC50 of luteolin on C918 cells for 24 h and 48 h were 51.46 μmol/L and 28.56 μmol/L,respectively;the IC50 of luteolin on OCM-1 cells for 24 h and 48 h were 54.17 μmol/L and 31.39 μmol/L,respectively.The results of plate clone formation assay and EDU cell proliferation assay showed that luteolin could effectively inhibit the proliferation and clone formation of C918 cells and OCM-1 cells,and all showed significant dose dependence.Hoechst staining and Annexin Ⅴ-FITC/PI double staining flow cytometry revealed that luteolin can also induce apoptosis in C918 cells and OCM-1 cells.Matrigel cell adhesion assay,wound scratch assay,Transwell cell migration assay and Transwell cell invasion assay showed that luteolin can effectively inhibit the adhesion,migration,and invasion of C918 cells and OCM-1 cells,and the more significant the inhibition effect with the increase of drug dose.ELISA showed that luteolin could inhibit MMP-2 secretion of C918 cells and OCM-1 cells.Western blot showed that the expression of vimentin in C918 cells and OCM-1 cells was not affected when luteolin concentration was 15μmol/L,but decreased when luteolin concentration was 20 μmol/L.Western blot also showed that luteolin reduced the expression of p-PI3 K P85 and p-Akt in C918 cells and OCM-1 cells.After IGF-1 pretreated C918 cells and OCM-1 cells,luteolin was added.The results showed that there was no significant difference in the number of cell clone formation between the group of 15 μmol/L luteolin + IGF-1 and the group of 15 μmol/L luteolin(P > 0.05);The results of Matrigel cell adhesion assay showed that the inhibition of luteolin on the adhesion of OCM-1 cells was weakened after IGF-1 pretreatment,but in C918 cells,there was no significant difference between 15μmol/L luteolin + IGF-1 group and 15 μmol/L luteolin group(P > 0.05);Transwell cell migration assay and Transwell cell invasion assay showed that luteolin inhibited migration and invasion of C918 cells and OCM-1 cells after IGF-1 pretreatment.Conclusion:Luteolin inhibited the proliferation,adhesion,migration,invasion and induced apoptosis of C918 cells and OCM-1 cells.Luteolin can inhibit the secretion of MMP-2 in C918 cells and OCM-1 cells,and inhibit the expression of cytoskeleton protein Vimentin.Luteolin inhibits the migration and invasion of C918 cells,and the migration and invasion and adhesion of OCM-1 cells are related to the inhibition of the PI3K/Akt signaling pathway;Luteolin may inhibit C918 cell adhesion,C918 cell clone formation,and OCM-1 cell clone formation through other signaling pathways.