Experimental Study On The Effects Of Curcumin On The Mechanisms Of Proliferation And Apoptosis In The Human Chordoma CM-319Cell Lines

ObjectiveTo study the experimental study on the effects of curcumin on the mechanisms ofproliferation and apoptosis in the Human chordoma CM-319cell lines. To discuse thefunction of curcumin on chordoma CM-319cell and its possible mechanism ofproliferation and apoptosis.MethodsPeople chordoma CM-319cells were training in vitro, CM-319cells were treatedwith different concentrations of curcumin(0μmol/L、10μmol/L、20μmol/L、30μmol/L、40μmol/L、0.1％DMSO) and for different hours（12h、24h、48h）， CCK-8wasused to determine growth inhibition. CM-319cells were treated with differentconcentrations of curcumin(0μmol/L、10μmol/L、20μmol/L、30μmol/L、40μmol/L) for24hours, collected and fixed cells, Flowcytometry was used to examineapoptosis and analysed Cell cycle. CM-319cells were treated with differentconcentrations of curcumin(0μmol/L、20μmol/L、40μmol/L) and for different hours（0h、24h、48h），collectiing cells, cracking cells on ice，extracting and quantitatingprotein. Caspase-3,Caspase-8, Caspase-9relative activities were tested bycolorimetry.Results1、 CCK-8test results:It showed that the curcumin had the function asproliferation and inhibition on CM-319cells, and it was line-depended on theconcentration and time， and the differences were great(p<0.05).The highestproliferation inhibition rate is: the application of40μmol/L of curcumin treatmentafter48hours,and the CM-319cells proliferation inhibition rate reached64.3%.Littleinfluence is found on the growth of cells treated with0.1%DMSO(p＞0.05).2、 FACS analysis indicated that the CM-319cells were effected by different concentrations of curcumin (0μmol/L,10μmol/L,20μmol/L,30μmol/L,40μmol/L)after24h, the cycle distribution has changed significantly.With the increase of drugconcentration, the number of G1phase cells were increased gradually,and the numberof S and G2phase cells were reduced gradually. Each curcumin group had asignificant difference compared with control group (p <0.05). After CM-319cellswere exposed to curcumin(10μmol/L、20μmol/L、30μmol/Land40μmol/L) for24h,the apoptosis rates were9.27％、11.70％、17.56％、21.89％respectively,whichwere significantly higher than control2.7％(p＜0.05).3、The CM-319cells were effected by different concentrations of curcumin(0μmol/L、20μmol/L、40μmol/L) after different time points（0h、24h、48h）： theactivities of Caspase-3were significantly increased with the extension of time and theconcentration of curcumin, each curcumin group had a significant differencecompared with control group and the same concentration in different time period(p <0.01); the activities of Caspase-8and Caspase-9were significantly increased in12h,and reached its peak at24h, each curcumin group had a significant differencecompared with control group and the same concentration in different time period(p <0.01);Conclusion1、The curcumin can significantly inhibit the proliferation of chordoma CM-319cells in vitro, and it also can induce chordoma CM-319cells apoptosis occurred andcause a cell cycle block.2、Curcumin induces apoptosis of human chordoma CM-319cells maybe byactivating Caspase-8、 Caspase-9, and then through different pathways risingCaspase-3activity.