Studay On The Inhabitation And Related Apoptosis Factor Of ACC-M Cells Induced By Allicin Combined With 5-FU

Objective:To investigate the apoptosis effect of allicin combined 5-fluorouracil on adenoid cystic carcinoma?ACC-M?cells,and to explore its possible mechanisms through regulating the expression of Bcl-2 and caspase-3 related proteins.This paper provides a certain theoretical basis and experimental basis for the treatment of adenoid cystic carcinoma with the combination of allicin and 5-fluorouracil.Experimental design:?1?The allicin group,5-fluorouracil group and allicin+5-fluorouracil group with concentration gradient?25,50,75,100,125?g/ml?were used to incubate ACC-M cells together with the control group.After 12,24,and 36 hours,the CCK-8 method was used to detect the inhibition rate of tumor cells in each concentration group after different time.The experimental results were statistically analyzed,and the median lethal concentration of allicin and 5-fluorouracil on tumor cells in 24 hours was calculated for subsequent experiments.?2?After data collection and analysis,the ACC-M cells?12,24h?were cultured with allicin,5-fluorouracil and allicin+5-fluorouracil at concentrations of?25,50,75,100,125?g/ml?.The apoptosis was detected by flow cytometry,and the result data was statistically analyzed.?3?Through the calculation of IC50 value,the ACC-M cells were separately and co-cultured with allicin with a concentration of 75.00?g/ml and 5-fluorouracil with a concentration of 68.50?g/ml;Western blot was used to detect changes in Bcl-2 protein and caspase-3 protein associated with classical apoptosis 24 hours after administration.Results:?1?The comparison between the concentration of each drug in the experimental group and the control group on the proliferation inhibition of ACC-M cells was statistically significant?F=342.195,P<0.01?.The inhibitory effect of allicin on the proliferation of adenoid cystic carcinoma cells gradually increased with the increase of drug action time?F=79.83,P<0.01?.?2?As the drug concentration increased,the 5-fluorouracil group had an increasing inhibitory effect on adenoid cystic carcinoma cells.?F=483.44,P<0.01?.?3?When 75.00?g/ml allicin combined with 68.50?g/ml 5-fluorouracil group acted on ACC-M cells,its inhibitory effect on tumor cell proliferation was stronger than that of the two used seperately in the 12h period,and the difference was statistically significant?F=210.18,p<0.01?;?4?Flow cytometry's detection showed that each experimental group induced apoptosis of ACC-M cells to varying degrees,and the apoptotic rate increased with the increase of drug concentration and the duration of action.At 12h and 24h period,the apoptotic rate of ACC-M cells in high,medium and low concentrations was:allicin group<5-fluorouracil group<allicin+5-fluorouracil group?P<0.05?;The difference between the combination group and each individual drug group was statistically significant.?P<0.05?,indicating that the combination group had the strongest killing effect on ACC-M cells.?5?.5Western bolt detection:using allicin and 5-fluorouracil separately or in combination can decrease the expression of Bcl-2 protein and increase the expression of caspase-3protein.The allicin group has a slightly weaker ability to regulate related proteins compared with the 5-fluorouracil group?P<0.05?,and the combination of the two drugs has a better effect to regulate the target protein than the drug alone?F₁=159.00,P<0.01??F₂=594.20,P<0.01?.Conclusion:?1?Allicin and 5-fluorouracil at different concentrations had the ability to inhibit proliferation and induce apoptosis of ACC-M cells.Compared with the single component experimental group,the experimental group with two components used together had stronger effect on inhibiting proliferation and inducing apoptosis of ACC-M cells.?2?Allicin combined with 5-fluorouracil may inhibit proliferation and induce apoptosis of ACC-M cells by inhibiting the activity of Bcl-2 and up-regulating the expression of caspase-3.