Synergistic Effect And Mechanism Of S-allylcysteine Combined With 5-fu On Proliferation Inhibition Of Hepatocellular Carcinoma HEP G2 Cells

Primary liver cancer is a common malignant tumor in China,and the 5-year recurrence rate of patients undergoing radical resection is about 70%.The important reasons for poor prognosis in patients with liver cancer include recurrence after treatment.Hepatocellular carcinoma is less sensitive to most anticancer drugs,with most therapeutic effects of about 10 to 20 percent.Therefore,it is of great research significance and practical value to find a drug with good curative effect and low toxicity or a drug that can enhance the curative effect of chemotherapy drugs and reduce the dosage.S-allylcysteine is one of the main active components in garlic extract.In recent years,studies have shown that S-allylcysteine has significant tumor inhibition effect.In this paper,the effects of SAC combined with 5-Fu on the proliferation ability of HEP G2 cells and the related molecular mechanisms were studied through in vitro experiments.Purpose:To investigate the inhibitory effect of S-allylcysteine and 5-FU on the proliferation of hepatocellular carcinoma HEP G2 cells and its molecular mechanism.Methods:1.Study on the effect on the survival rate of HEP G2 cells:hepatoma cells were cultured in vitro,and the effect of SAC and 5-FU alone or in combination on the survival rate of HEP G2 cells was confirmed by CCK8 experiment.2.Study on the effect of cell apoptosis:The effect of SAC and 5-FU alone or in combination on the existence and promotion of apoptosis of HEP G2 cells was observed by flow cytometry and under inverted microscope.3.Western blot assay was used to detect the changes in the expression levels of bcl-2 and bcl-xl proteins in HEP G2 cells after SAC combined with 5-FU or single treatment.Results:1.In In the experiment of the inhibition of HEP G2 cell proliferation by SAC and 5-FU alone and in combination with SAC and 5-FU:(1)Compared with the control group,the survival rate of HEP G2 cells in SAC and 5-FU alone and in combination with SAC and 5-FU alone and in combination with SAC and 5-FU alone and in combination with SAC and 5-FU was lower,and the effect was more obvious with the increase of drug concentration.(2)The survival rate of HEP G2 cells with SAC and 5-FU was lower than that with SAC and 5-FU alone.(3)According to the medium-effect principle and formula,the medium-effect concentration of SAC and 5-Fu alone was calculated to be 289.14umol/L and 20.571umol/L,respectively.When 41umol/1SAC was combined with 5-Fu,the medium effective concentration of 5-Fu was 10.798umol/L.By calculating the joint index CI= 0.667&It when the cell inhibition rate was 50%;1.It is suggested that SAC combined with 5-FU has a synergistic effect on the proliferation inhibition of HEP G2 cells.2.The morphology of HEP G2 cells was observed under inverted phase contrast microscope.Compared with SAC and 5-FU alone group,the floating dead cells in SAC combined with 5-FU group were significantly increased,indicating that the combined group had a stronger inhibitory effect on proliferation.3.Flow cytometry showed that the apoptosis rate of SAC and 5-FU combined group was 50.8%,and that of 5-FU progenitor cells was 30.75%.The apoptosis rate of SAC group was 16.563%.The apoptosis rate of the control group was 2.453%.These results indicated that the combination group and single group could inhibit the proliferation of HEP G2 cells by inducing apoptosis.4.Western blot results showed that SAC combined with 5-Fu and single use could reduce the expression levels of Bcl-2 and Bcl-XL proteins.The gray values of Bcl-2 and Bcl-xL in the control group were 1.435±0.311 and 1.789±0.336,respectively.The gray values of Bcl-2 and Bcl-xL in SAC group were 1±0.206 and 1.214±0.203,respectively.The gray values of Bcl-2 and Bcl-xL in 5-Fu group were 0.599±0.191 and 0.894±0.103,respectively.The gray values of Bcl-2 and Bcl-xL in SAC and 5-FU groups were 0.279±0.105 and 0.537±0.041,respectively.The results showed that SAC combined with 5-Fu reduced the expression of Bcl-2 and Bcl-xL in HEP G2 cells more obviously than the single treatment group.Conclusion:1.SAC has a synergistic effect on 5-Fu induced proliferation inhibition of HEP G2 cells.2.SAC combined with 5-Fu induced apoptosis of HEP G2 cells,which inhibited cell proliferation.3.SAC combined with 5-Fu can reduce the expression of Bcl-X and Bcl-2 proteins in HEP G2 cells more significantly than single use,so as to exert synergistic inhibition.