Cell-killing and sensitization effect of 6-gingerol on human hepatoma carcinoma cell in chemotherapy in different environment / 中华老年医学杂志

ABSTRACT

Objective To compare the cell-killing and sensitization effect of 6-gingerol on human hepatoma carcinoma (HepG-2) cell in normal mode versus hypoxia-hypoglycemia mode in chemotherapy.Methods The HepG-2 cells was cultured to logarithmic phase and treated with adriamyein doxorubicin hydrochloride (ADM) (5,10,15,20,40,60 mg/L) and 6-gingerol(25,50,100,200 μmol/L)in different concentrations.Then the cell counting kit (CCK-8) assay kit was used to determine the proliferation inhibition of HepG-2 cells.Cell apoptosis was detected by combining flow cytometry and AnnexinV-FITC PI double staining after treated with different drugs.The expressions of bcl-2,bax and birc-5 mRNA in HepG-2 cells was detected by real time fluorescence quantitative polymerase chain reaction(RT-PCR)assay.Results 6-gingerol and ADM had a certain degree of growth inhibition on HepG-2 cells.In two modes,the inhibition ratios of the 6-gingerol and ADM were both increased along with the increase of the concentration,which showed a dose-dependent manner.Flow cytometry analysis demonstrated that the apoptosis rate in the control group,6-gingerol group,ADM group and the 6-gingerol+ADM group in the normal mode was (7.98±0.76)％,(9.63 ± 1.00) ％,(12.70 ± 2.13) ％ and (19.92 ± 1.41) ％ respectively.The apoptosis rate in the control group,6-gingerol group,ADM group and the 6-gingerol+ ADM group in the hypoxia-hypoglycemia mode was (13.92 ± 2.02)％,(19.36 ±-1.22)％,(27.87 ± 0.99)％ and (38.63 ± 2.25)％ respectively.It demonstrated that the apoptosis rate was increased in the experimental groups as compared to the control group under the two culture conditions(the normal mode and the hypoxiahypoglycemia mode)(t=7.250,5.259,12.185,8.140,15.000,47.576,respectively,all P＜0.05,0.01 or 0.001).The combination group had the highest number of apoptosis cells,and the number of apoptosis cells was higher in hypoxia-hypoglycemia group than in normal culture group.Real-time PCR analysis showed that,compared with the control group,the expressions of bcl-2 and birc-5 mRNA were decreased and the expression of bax mRNA had no significant changes in experimental group under the normal culture conditions.The expressions of bcl-2 and birc-5 mRNA were significantly decreased and the expression of bax mRNA was increased in experimental group as compared with the control group under the hypoxia-hypoglycemia conditions.Under the hypoxiahypoglycemia environment,the expression of bcl-2 mRNA was significantly increased,the expressions of bax and birc-5 mRNA was significantly reduced,and the ratio of bcl-2 and bax was significantly increased as compared with the normal culture conditions.Conclusions 6 gingerol may decrease the inhibitory effect of survivin protein on tumor cells apoptosis by reduced the expression of birc-5,which generates the cell-killing and sensitizing effect on HepG-2 cell in chemotherapy.This performance is more obvious in the hypoxia-hypoglycemia environment.