Effect and mechanism of curcumin on proliferation and apoptosis of human hepatoma HCC-LM3 cells / 国际外科学杂志

ABSTRACT

Objective To investigate the effects of curcumin on the growth and apoptosis of human hepatoma HCC-LM3 cells and explore the molecular mechanisms.Methods The human hepatoma HCC-LM3 cells were treated with different concentrations of curcumin and the cell growth inhibition rate was detected by CCK-8.The effect of curcumin on human hepatoma HCC-LM3 cells was observed.Refer to the relevant literature,the human hepatoma HCC-LM3 cells were treated with the concentration of 2.5,5.0,10.0,15.0,20.0,40.0,60.0 μmol/L of curcumin for 48 hours,taking the 0 μmol/L curcumin as control group,and the cell growth inhibition rate was detected by CCK-8.According to the results of CCK-8,selecting the concentration of 0 μmol/L as control group and the concentration of 10.0,20.0,40.0 μmol/L as experimental groups,which has significant difference on growth inhibition rates.Cell cloning assay was used to detect cell cloning ability,Flow cytometry was used to detect apoptosis,and Western blotting to detect the protein expression levels of Mcl-1,Bax,Bcl-2 and Bcl-xL.The measurement data were expressed in ((x) ± s),and the single factor analysis of variance was used for comparison between groups.Results CCK-8 assay showed that with treated by the concentration of 2.5,5.0,10.0,15.0,20.0,40.0,60.0 μmol/L,the growth inhibition rates were(6.71 ± 3.45)％,(12.33 ± 5.02)％,(20.07 ± 5.60)％,(57.80 ±7.34)％,(78.37 ±6.53)％,(91.73 ±6.14)％ and (96.18 ±3.45)％,suggesting that curcumin could inhibit the growth of human hepatoma HCC-LM3 cells in a dose-dependent manner.Cell clone formation experiment showed that curcumin could inhibit the clone of the human hepatoma HCC-LM3 cells,and the clone of the cells was inhibited significantly when the concentration of the curcumin was over 20.0μmol/L.The result of Annexin V-FITC/PI double staining analysis showed that the apoptotic rates of experimental groups and control groups were (5.20 ± 1.44) ％,(9.90 ± 3.31) ％,(55.67 ± 5.29) ％,(79.63 ±4.71)％,with all the apoptotic rates of experimental group over the control groups (P ＜0.05),suggesting curcumin could induce the apoptosis of human hepatoma HCC-LM3 cells.The Westen blotting showed that curcumin increased the expression of Bax protein while decreasing expression of Mcl-1 protein significantly in concentration-dependent manner (P ＜ 0.05),but have no effect on the expression of Bcl-2 and Bcl-xL proteins.Conclusion Curcumin could inhibit the proliferation and clone of human hepatoma HCC-LM3 cells,and induce apoptosis in a dose dependent manner.