Abstract
Objective To explore the effects of curcumin on the in vitro proliferation and apoptosis of mouse liver can-cer Hepa1-6 cells.Methods Hepa1-6 cells in the logarithmic growth phase were divided into the control group,10 μmol/L group,20 μmol/L group and 40 μmol/L group,respectively,which were treated with 0,10,20,and 40 μmol/L of cur-cumin.Cell proliferation was assessed by the MTT assay,and apoptosis level was detected by FACS.Real-time quantitative PCR(RT-qPCR)was employed to detect the levels of apoptosis-related molecules Bcl-2,Caspase-3,Toll-like receptor 2(TLR2),IFN-α,and IFN-β in Hepa1-6 cells.The secretion levels of IFN-α and IFN-β in the cell culture supernatant were determined using ELISA.Hepa1-6 cells were incubated with CD4+T cells,CD8+T cells and NK cells in the control group and 40 μmol/L curcumin group,respectively.FACS was used to detect the expression levels of CD69 in CD4+T cells,CD8+T cells and NK cells.Results The proliferation inhibition rates of Hepa1-6 cells in the 10,20 and 40 μmol/L groups increased in a dose-dependent manner,and the inhibition rates were(36.45±2.11)%,(46.40±2.08)%and(82.16±3.32)%,respectively.There was statistically significant difference in comparison with that of the control group(P<0.05).The apoptotic proportions of Hepa1-6 cells in the 10 μmol/L,20 μmol/L and 40 μmol/L groups were 10.47%,26.66%and 49.81%,respectively.And the difference was statistically significant in comparison with that of the control group(P<0.01).RT-qPCR results showed that the transcription levels of anti-apoptotic gene Bcl-2 in the 10 μmol/L,20 μmol/L and 40 μmol/L groups were(0.77±0.06)times,(0.44±0.06)times and(0.25±0.08)times that of the control group,respectively,with statistically significant difference(all P<0.01).At the same time,the expression of pro-apoptotic gene Caspase-3 significantly increased,and the transcription levels of pro-apoptotic gene Caspase-3 in the 10 μmol/L,20 μmol/L and 40 μmol/L groups was(1.73±0.31)times,(2.52±0.45)times and(3.25±0.20)times that of the control group,respectively,with statistically significant difference(all P<0.01).With the increase of curcumin concentrations,TLR2 transcription levels increased gradually.TLR2 transcription levels in the 10 μmol/L,20 μmol/L and 40 μmol/L groups were(3.56±0.59)times,(4.93±0.04)times and(6.60±0.47)times that of the control group,respectively,with statistically significant difference(all P<0.05).In addition,the transcription levels of IFN-α in the 10 μmol/L,20 μmol/L and 40 μmol/L groups significantly increased,which were(4.21±0.22),(5.54±0.39)and(8.05±0.93)times that of the control group,respectively,and there were statistically significant differences among these groups(all P<0.05).The transcription levels of IFN-β were(3.08±0.51),(5.31±0.55)and(8.35±0.59)times that of the control group,and the difference was statistically significant(all P<0.01).For IFN-α,compared with the control group[(36.26±6.12)pg/mL],the levels of IFN-α secreted by cells in the 10 μmol/L,20 μmol/L and 40 μmol/L groups were(57.00±8.8),(82.47±4.42),and(100.37±8.99)pg/mL,respectively;the levels of IFN-α secreted by cells in the 10,20,and 40 μmol/L groups were higher that that of the control group(all P<0.01).The levels of IFN-β secreted by the cells in the 10,20,and 40 μmol/L groups were(60.67±6.81),(91.33±5.51),and(144.00±10.15)pg/mL,respec-tively,while that in the control group was(44.67±1.15)pg/mL.The levels of IFN-β secreted by the cells in the 10,20,and 40 μmol/L groups were higher than that in the control group(all P<0.01).In the control group,CD69 expression lev-els of CD4+T cells,CD8+T cells and NK cells were(24.53±1.39)%,(23.40±2.32)%,(32.33±3.08)%,respectively.The CD69 expression levels of CD4+T cells,CD8+T cells and NK cells in the 40 μmol/L group were(39.90±3.51)%,(31.47±3.88)%,(50.83±4.74)%,respectively.There were significant differences in CD69 levels of CD4+T cells,CD8+T cells and NK cells between control group and 40 μmol/L group(all P<0.05).Conclusion Curcumin promotes the secretion of type Ⅰ interferon by regulating the expression of TLR2,thus inhibits the proliferation of hepatocellular car-cinoma cells,induces the apoptosis of hepatocellular carcinoma cells,as well as promotes the activation of immune cells.关键词
肝癌/姜黄素/Toll样受体2/细胞增殖/细胞凋亡/免疫功能Key words
hepatocellular carcinoma/curcumin/Toll-like receptor 2/cell proliferation/apoptosis/immune function分类
医药卫生
