Effect Of Disulfiram Combind With Copper Ion On Proliferation Of Hela Cells In Cervical Cancer

Objective:To study the effects of different concentrations of Disulfiram?DSF?and Disulfiram combined with Cu ion?DSF/Cu?on the proliferation of human cervical cancer Hela cells in vitro,and to explore its relationship with NF-?B and PI3K/Akt/mTOR protein pathway.Methods:1.The Hela cell line of human cervical cancer was used as the research object,and Cu?1?mol/L?,different concentrations of DSF?0.1?mol/L,0.2?mol/L,0.4?mol/L,0.8?mol/L,1.6?mol/L,3.2?mol/L,6.4?mol/L?and DSF/Cu?The concentration of 0.1?mol/L,0.2?mol/L,0.4?mol/L,0.8?mol/L,1.6?mol/L,3.2?mol/L,6.4?mol/L DSF combined with 1?mol/L Cu?were applied to human cervical cancer Hela cells for 24 hours,and 5 replicate wells were set for each concentration gradient.The proliferation inhibition effect of Hela cells was detected by thiazole blue colorimetric assay?MTT assay?.The corresponding IC₅₀0 values were calculated by graph prims5 software.2.Western blotq?WB?was used to detect the expression of p65,PI3K,Akt,p-Akt and mTOR protein in each group of cells treated with Cu,DSF and DSF/Cu.3.Statistical analysis was performed using SPSS19.0 statistical software.The experimental data of each group were expressed by mean±standard deviation?x±s?.The comparison of the mean of two independent samples was performed by t test.The comparison of multiple sample means was used to select one-way variance.Analysis,the test standard is?=0.05.Results:The MTT assay showed that the inhibition rate of Cu?1?mol/L?was?1.00±0.6?%,and there was no significant difference compared with the control group?P>0.05?.The cell proliferation inhibition rates of DSF with 0.1?mol/L,0.2?mol/L,0.4?mol/L,0.8?mol/L,1.6?mol/L,3.2?mol/L,or 6.4?mol/LDSF for 24h were12.08%,12.87%,13.11%,14.74%,15.66%,16.47%,18.98%,compared with the control group were statistically significant?P<0.0001?and concentration-dependent?F=2.692,P<0.05?.Treated with different concentrations of DSF combined with copper ions?1?mol/L?,the cell proliferation inhibition rate of human cervical cancer Hela cells increased significantly,which were 30.13%,60.23%,71.65%,87.75%,91.23%,94.88%,97.66%,respectively.Compared with the control group,the difference was statistically significant?P<0.0001?,and the inhibition increased with the increasing of concentration?F=215.7,P<0.0001?.The inhibitory effect of DSF/Cu on the proliferation of Hela cells was more obvious than that of DSF alone.The IC₅₀ of the two groups were?2.460±0.008??mol/L and?0.053±0.010??mol/L.Western blot showed that the expression levels of p65 and p-Akt protein in DSF and DSF/Cu Hela cells decreased?P<0.05?,but no significant effect on the expression of PI3K,Akt and mTOR proteins?P>0.05?,and the p65 and p-Akt proteins in the DSF/Cu-treated group were more significant than those in the DSF group.Conclusion:1.DSF/Cu and DSF can inhibit the proliferation of Hela cells,and the effect of DSF combined with Cu ion is significantly stronger than that of DSF alone.2.Its mechanism may be related to inhibition of NF-?B and PI3K/Akt/mTOR protein pathways.