Curcumin Mediates Apoptosis,migration And Invasion In Giant Cell Tumor Of Bone By Targeting JAK2/STAT3 Signal Pathway

BackgroundGiant cell tumor of bone(GCTB)is one of the common primary bone tumors and considered as the non-aggressive or low potential tumor,with regional invasion and usually involving the ends of the long bones.Curcumin was the nature phenolic antioxidant O For the anti-tumor of curcumin,clinically,curcumin is taken as the main component for prevention and treatment on different kinds of tumor.At present,the therapeutic effect of curcumin in giant cell tumor of bone has not been reported.ObjectiveIn this study,we investigated the effects of Curcumin on the biological behavior of Giant cell tumor of bone through experiments.The aim of this study was to elucidate the anti-Giant cell tumor of bone mechanism of Curcumin.Method1.The clinical specimens were collected and collected for primary cell culture.The effects of curcumin on proliferation,apoptosis,migration and invasion of GCTB cells were investigated by CCK8,flow cytometry and Transwell to identify the biological function of curcumin on GCTB.2.Setting groups:A.NC(0.1%DMSO)group;B.Cur(31.5nM)group;C.AG490(1μM)group;D.STAT3 siRNA(100nM)group;E.JAK2 siRNA(100nM)group;F.Cur(31.5nM)+ STAT3 siRNA(100nM)group;G.Cur(31.5nM)+ JAK2 siRNA(100nM)group;H.Cur(31.5nM)+ AG490(1μM)group.Western Blot(WB)detection method was used to detect the effect of curcumin on the expression level and activation state of JAK2/STAT3 signaling pathway protein in GCTB cells.3.By using STAT3,siRNA alone or together with JAK2 siRNA or JAK2/STAT3 signal pathway blocker AG490,observe the effects of curcumin on promoting GCTB cell apoptosis and inhibit GCTB cell migration and invasion activity,biological effect clear whether curcumin by acting on JAK2/STAT3 signal pathway play anti GCTB effect.Setting groups:A.NC(0.1%DMSO)group;B.Cur(31.5nM)group;C.AG490(1μM)group;D.STAT3 siRNA(100nM)group;E.JAK2 siRNA(100nM)group;F.Cur(31.5nM)+ STAT3 siRNA(100nM)group;G.Cur(31.5nM+ JAK2 siRNA(100nM)group;H.Cur(31.5nM)+ AG490(1μM)group.Flow cytometry,Transwell and other experimental methods were used to detect the changes of cell apoptosis and migration,invasion detection and so on.Results1.31.5nM curcumin was taken as the finally treating concentration,which could inhibit GCTB cell proliferation stably;curcumin(31.5nM)treating for 48h could induce 50%of the GCTB cells apoptosis;curcumin(31.5nM)treating for 48h could inhibit GCTB cells migration and invasion.2.The result as follows:(1)31.5nM curcumin can inhibit the phosphorylation of STAT3 protein alone.(2)31.5nM curcumin can inhibit the phosphorylation of JAK2 protein alone;(3)31.5nM curcumin can simultaneously inhibit the phosphorylation of STAT3 and JAK2 protein;(4)31.5nM curcumin and JAK2/STAT3 signaling pathway blocker AG490(1 mu M)jointly inhibit the conduction of JAK2/STAT3 signaling pathway.3.The results showed that STAT3 and JAK2 can promote the migration and invasion of GCTB cells;JAK2/STAT3 signal pathway blocking agent AG490 can inhibit the migration and invasion of GCTB cells;curcumin by acting on JAK2/STAT3 signaling pathway to inhibit the migration and invasion of GCTB cells;synergistic inhibitory effect of curcumin can increased JAK2/STAT3 signal pathway inhibitor AG490 on migration and invasion of GCTB cells.Conclusioncurcumin by acting on JAK2/STAT3 signaling pathway to could induce of the GCTB cells apoptosis,inhibit the migration and invasion of GCTB cells.