Effect Of Oxysophocarpine On Proliferation And Apoptosis Of Hepatoma Cells

Objective:Oxysophocarpine is an effective component extracted from Sophora flavescens and belongs to dehydrogenated matrine alkaloids.With the further study of matrine alkaloids in modern pharmacology,some studies have found that matrine alkaloids have anti-viral,anti-inflammatory and anti-tumor effects.It has been confirmed that oxysophocarpine can inhibit the proliferation and induce apoptosis of colon cancer SW480 cells and breast cancer MCF-7 cells in vitro.At present,there is still no effective treatment for hepatocellular carcinoma,and the effect of oxysophocarpine on hepatocellular carcinoma cells has not been reported.In this study,oxysophocarpine and SMMC-7721 cells were studied.Inquiry:1.Effects of sophoridine oxide on cell proliferation and apoptosis;2.The specific action mechanism of sophoridine oxide on hepatocellular carcinoma cells provides a theoretical basis for the clinical application of drugs.Methods:?1?SMMC-7721 cells and L-02 cells were treated with oxysophocarpine at a concentration gradient and cultured for 48 hours after drug treatment.The morphological changes of SMMC-7721 cells were observed by inverted microscope.?2?CCK-8 method was used to detect the effects of oxysophocarpine with different concentration gradients on the proliferation of SMMC-7721 cells and L-02 cells,and the appropriate concentration of oxysophocarpine was screened after calculating IC₅₀.?3?Flow cytometry was used to detect the effect of oxysophocarpine at different concentrations on apoptosis of SMMC-7721 cells.?4?Western blot was used to detect the expression of Bcl-2,Bax,STAT3 and STAT5in SMMC-7721 cells.Results:?1?Morphological changes of hepatocellular carcinoma cells 48 hours after treatment.Compared with the control group,SMMC-7721 cells proliferation was inhibited with the increasing concentration of oxysophocarpine 2,4,8 mmol/L,and some cells shrank and died.At the concentration of 16 mmol/L,the number of hepatocellular carcinoma cells in the microscopic field of vision was significantly less than that in the control group,with a certain degree of cell death.At the concentration of 32 mmol/L,SMMC-7721 cell proliferation was significantly inhibited and most of the cells died.Compared with the control group,the number of L-02 cells in the visual field decreased slightly when the low concentration of drugs was applied to L-02 cells,while the number of L-02 cells decreased significantly when the concentration of sophocarpine was 32 mmol/L,and the cells shrank and died.?2?CCK8 assay showed that oxysophocarpine significantly inhibited the proliferation of SMMC-7721 cells compared with the control group,and OD value decreased significantly with the increase of concentration?P<0.05?.When the same concentration of oxysophocarpine acted on L-02 cells,there was no significant difference between the drug treatment group and the control group?P>0.05?,and the IC₅₀0 of oxysophocarpine acted on hepatocellular carcinoma cells was 5.294.?3?Flow cytometry showed that the apoptotic rate of SMMC-7721 cells treated with oxysophocarpine?2,4,8,16 mmol/L?for 48 hours was significantly higher than that of the other four groups?all P<0.05?,suggesting that oxysophocarpine could promote the apoptosis of SMMC-7721 cells.?4?Western blot analysis showed that the expression of anti-apoptotic protein Bcl-2was significantly decreased in oxysophocarpine-treated group compared with the control group?P<0.05?,while the expression of Bax was significantly increased in oxysophocarpine-treated group?P<0.05?.The expression of STAT3 and STAT5 in oxysophocarpine-treated group was significantly lower than that in control group?P<0.05?.Conclusions:Oxysophocarpine can inhibit proliferation and promote apoptosis of hepatocellular carcinoma cells.This may be related to down-regulation of STAT3 and STAT5 protein kinase expression through JAK-STAT signaling pathway,thus up-regulation of Bax gene expression and down-regulation of anti-apoptotic Bcl-2 gene expression.