Study On The Apoptosis And Its Related Mechanisms Of Mucoepidermoid Cancer Cell Line MEC -1 Induced By Allicin Combined With 5-FU

Objective:To study the apoptosis mechanism of MEC-1 cells induced by allicin combined with 5-FU and its effect on the protein expression of Bax and Bcl-2 involved in the apoptosis pathway,and to provide the basis for subsequent experiments and clinical drug application.Methods:Control group was treated with 0.1%DMSO in PBS,and the experimental group was allicin group,5-FU group,allicin combined with 5-FU group.The logarithmic growth phase cells were taken for experiments.（1）CCK8 assay was used to detect the proliferation inhibition rate of MEC-1 cells after different concentrations（0、25、50、75、100mg/L）of allicin,5-FU and allicin combined with 5-FU for a certain period of time（24、48、72h）.（2）Flow cytometry was used to detect the apoptotic rate of MEC-1 cells treated with allicin,5-FU and 5-FU at different concentrations（0、25、50、75mg/L）for a certain time（12、24h）.（3）After the IC50 concentration of allicin and IC50 concentration of 5-fu acted on mec-1 cells alone or in combination for 24 hours,the expressions of Bax and Bcl-2 proteins were determined using Westernbolt.Results:（1）At the same time,with the increase of drug concentration,the inhibition rate of MEC-1 cell activity in each experimental group increased.The inhibition rates of allicin group and 5-FU group were significantly different from those of combined drug group(P_group<0.01).（2）The growth trend of inhibition rate of MEC-1 proliferation in allicin group,5-FU group and combination group increased with the increase of time or concentration,and the inhibition rate difference between combination group and single group was statistically significant(P_group<0.01).（3）Compared with the control group,all three drugs can induce MEC-1 apoptosis.In addition,the intensity of apoptosis induced by MEC-1 cells in the three experimental groups increased with the increase of concentration and time.The apoptosis rate of MEC-1 cells in the combination group was significantly different from that in the allicin group and the 5-FU group(P_group<0.05),and the 75mg/L combination group showed the strongest apoptosis effect.（4）Allicin and 5-FU alone and in combination can down-regulate Bcl-2 protein and up-regulate Bax protein expression,and the ratio of Bax/Bcl-2 is the largest in the combination group（P<0.05）.Conclusion:（1）Both allicin and 5-FU,used alone or in combination,inhibited the proliferation and induced apoptosis of MEC-1 cells,and the effect intensity increased with the increase of time or concentration.In addition,allicin enhanced the apoptosis effect of5-FU on MEC-1 cells.（2）The apoptosis mechanism of MEC-1 induced by allicin and 5-FU alone or in combination may be related to the apoptosis of mitochondrial pathway.