Effect of glycyrol combined with 5-FU on proliferation,apoptosis and chemosensitivity of colon cancer cells
Objective To investigate the effects and mechanism of glycyrol(GC)plus 5-FU on the proliferation,apoptosis and chemo-sensitivity of colon cancer cells.Methods The human colon cancer cell line HCT116 cells were treated with 0,2.5,5,10,15,20 μmol/L 5-FU and 0,5,10,20,30,40 μmol/L GC for 48 h,respectively,and the cell survival was calculated by crystal violet staining method to determine the optimal treatment concentration.The HCT116 cells were treated with 10 μmol/L 5-FU,20 μmol/L GC and 10 μmol/L 5-FU+20 μmol/L GC for 12,24,36,48,60 h,respectively,and the cell survival rate was calculated by crystal violet staining to determine the optimal treatment time.The HCT116 cells were routinely cultured(control group)or treated with 10 μmol/L 5-FU,20 μmol/L GC and 10 μmol/L 5-FU+20 μmol/L GC for 48 h,respectively.The cell viability was determined by crystal violet staining,the apoptosis was determined by flow cytometry,and the expression levels of cleaved polyadenosine diphosphate ribose poly-merase(cleaved-PARP),programmed death ligand 1(PD-L1),phosphorylated histone H3(p-Histone H3)and phosphorylated protein tyrosine kinase 2(p-JAK2)were detected by Western blot.The HCT116 cells were treated with different concentrations of 5-FU,GC,or in combination at the concentration ratio of 1:2 for 48 h,and the Chou-Talalay equation was used to calculate the combination index(CI)to assess the synergistic effect of 5-FU and GC.Results The optimal concentrations of 5-FU and GC to inhibit cell proliferation were 10 μmol/L and 20 μmol/L,respectively,and the optimal time for cell proliferation inhibition by the combined treatment of 5-FU and GC was 48 h.The results of crystal violet staining showed that compared with control group,the cell viability was significantly reduced in 5-FU+GC group(P＜0.01).Compared with 5-FU group and GC group,the cell survival rate was reduced in 5-FU+GC group,but there was no statistically significant difference.Flow cytometry results showed a significanly higher apoptosis rate in 5-FU+GC group than in control group(P＜0.01).The combination index(CI)values of all combinations were less than 1,and the combined treatment of GC and 5-FU had a synergistic effect.Western blot results showed that compared with control group,cleaved-PARP was upregulated in HCT116 cells in 5-FU+GC group(P＜0.05),PD-L1 and p-JAK2 expression were downregulated(P＜0.05),and p-Histone H3 expression was downregulated(P＜0.01).Compared with 5-FU group,cleaved-PARP expression was significantly upreg-ulated in HCT116 cells in5-FU+GC group(P＜0.05),while p-Histone H3 expression was significantly downregulated(P＜0.01).Conclusion Combination of glycyrol and 5-FU can inhibit the expressions of PD-L1 and p-Histone H3 by downregulating the phospho-rylation level of JAK2,inhibiting the proliferation,inducing apoptosis,and increasing the chemosensitivity of HCT116 cells.
glycyrolcolon cancer cellscell proliferationcell apoptosischemosensitivity
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