An Experimental Research On The Impact Of 20(S)-Ginsenoside Rg3 On The Sensitivity Of Oxaliplatin Mediated By Copper Transport Related Proteins In Hepatocarcinoma Cell

Objective:To investigate the effect of 20(S)-ginsenoside Rg3 on the sensitivity of oxaliplatin mediated by copper transport related proteins in hepatocarcinoma cells and the realated mechanism by cell experiments,aimed,to provide some references for improving the efficacy of oxaliplatin and optimizing the combination of traditional Chinese medicine and western medicine in the treatment of hepatocellular carcinoma.Method:The sensitivity of MHCC97H cells to 20(S)-ginsenoside Rg3 and oxaliplatin was measured by MTT assay,and which was calculated to select the suitable drug concentration.Then,the MTT assay was used to detect the sensitivity of oxaliplatin after combining wih low concentration 20(S)-ginsenoside Rg3 in the MHCC97H cells.Drugs combined effect was judged by calculating the q value which can be used to determine to conduct the later experiment.The expression of Ctrl,Atox1,ATP7A and ATP7B mRNA in MHCC97H cells were detected by the real-time fluorescence quantitative polymerase chain reaction(RT-PCR).The expression of Ctrl,Atox1,ATP7A and ATP7B protein expression were detected by Western blotting.Results:In the experiment of MTT assay,the results showed that 20(S)-ginsenoside Rg3 and oxaliplatin had significant inhibitory effects on MHCC97H cell proliferation(P<0.05),and the the inhibitory effect on MHCC97H cells proliferation exhibited dose and time dependency.The maximum non-toxic concentration of 20(S)-ginsenoside Rg3 wasl00p,M,which was used to combine with oxaliplatin following the significant increase of inhibition rate in MHCC97H cells(P<0.05).q value showed that the combined effect of oxaliplatin and low concentration of 20(S)-ginsenoside Rg3 was synergistic or additive.RT-PCR and Western blotting were used to detect the expression of Ctrl,Atox1,ATP7A and ATP7B in MHC97H cells treated with 100μM 20(S)-ginsenoside Rg3 combined with different concentrations of oxaliplatin.The results showed that:(1)Compared with the control group,the expression of Ctrl mRNA and protein in the oxaliplatin monotherapy group or oxaliplatin with 20(S)-ginsenoside Rg3 group all decreased(P<0.05)The expression of Ctrl mRNA and protein in the ginsenoside Rg3 group was not significantly changed,but the expression of Ctrl mRNA and protein in the combination group was higher than that of the oxaliplatin monotherapy group(P<0.05).(2)Compared with the control group,the expression of Aoxl mRNA and protein did not change significantly in the oxaliplatin monotherapy group,20(S)-ginsenoside Rg3 group and the combination group(P>0.05).(3)The mRNA and protein expression of ATP7A and ATP7B in the combination group were significantly lower than those in the oxaliplatin monotherapy group(P<0.05).However,the oxaliplatin group had no significantly change conmpared with the concrol group(P>0.05)and the combination group was lower than the control group(P<0.05).Conclusion:20(S)-ginsenoside Rg3 and oxaliplatin monotherapy can effectively inhibit the proliferation of MHCC97H cells.The low concentration of 20(S)-ginsenoside Rg3 monotherapy has small inhibitory effect in MHCC97H cell proliferation,but can significantly enhance the sensitivity of hepatocarcinoma cells to oxaliplatin for drug combination,in which the exist mechanism is a low concentration of 20(S)-ginsenoside Rg3 can prevent oxaliplatin from inducing Ctrl downregulation in a certain extent and can downregulate the expression of ATP7A and ATP7B.