Effect Of Sophoridine On Proliferation And Apoptosis Of Hepatocellular Carcinoma SMMC-7721 Cells In Vitro

Objective:To explore the effect of sophoridine on the proliferation and apoptosis of liver cancer SMMC-7721 cells in vitro and provide a new basis for drug treatment of liver cancer.Methods:(1)Cell culture: human liver cancer SMMC-7721 cells with DMEM containing 10% fetal bovine serum,the saturated humidity incubator at 37 ? and 5% CO2 in 2-3 d replacement medium,the logarithmic phase cells for subsequent experiments.(2)Cell treatment and experimental grouping: Take the cell,the pancreatic enzyme digestion after centrifugation,supernatant,after heavy hanging in the culture medium,adjust the cell density of about 1 * 10^4 / mL vaccination in 96-well plates,100 uL/ hole,in 37 ? and 5%CO2 incubator culture overnight to adherent cells thoroughly,and the cells were randomly divided into 6 groups,the control group cells were not treated,and the treatment group was divided into five groups according to the concentration of Sophoridine added.sophoridine concentrations were 0.5 mmol/ L,1 mmol/ L,2 mmol/ L,4 mmol/ L,8 mmol/ L.Set six parallel holes in each group.The morphological changes of cells were observed by inverted microscope after continuous culture for 48 h.(3)The effect of different concentrations of sophoridine on the proliferation of SMMC-7721 cells of liver cancer was detected by CCK8 experiment.(4)The effect of different concentrations of sophoridine on apoptosis of SMMC-7721 cells of liver cancer was detected by flow cytometry.(5)Differences in expression levels of apoptotic proteins Bcl-2 and Bax were detected by western blotting.Results:(1)When the concentration of sophoridine increased,SMMC-7721 cells were sparse and some cells appeared shrinkage and apoptosis.(2)The IC50 of SMMC-7721 treated with sophoridine is 7.167(95%CI: 5.167-9.943).(3)The results of CCK8 showed that the proliferation of sophoridine treated group(8 mmol/L)was significantly inhibited compared with the control group(P<0.05).(4)Flow cytometry showed that when the concentration of SMMC-7721 cells treated with sophoridine increased,the total apoptosis rate also increased.(P < 0.05).(5)Western blotting showed that with the increase of sophoridine concentration,Bax protein expression in SMMC-7721 cells was significantly higher than that in the other 5 groups(P<0.05),while Bcl-2 protein expression was significantly lower than that in the other 5 groups(P<0.05).Conclusion:Sophoridine may inhibit cell proliferation and promote apoptosis by up-regulating Bax expression and bcl-2 expression in SMMC-7721 cells of liver cancer.