| OBJECTIVE: To study the anti-hepatocellular activity and mechanism of mollugin,and to provide scientific basis for its development and application in the treatment of liver cancer.Methods: Human liver cancer Hep G2 cells were used as test subjects.MTT assay was used to detect the inhibitory effect of mollugin on Hep G2 cells at different concentrations and time.The plate cloning method was used to detect the long-term effect of mollugin on Hep G2 cells.Inhibition of proliferation.The mouse H22 liver cancer cell line was selected and BALB / c mice were injected subcutaneously to establish a mouse model of H22 hepatocellular carcinoma transplantation.Adriamycin was used as a positive control drug to observe the effect of mollugin on tumor growth,mouse growth status,and organs.At the same time,H&E staining was used to examine the changes of tumor cells in each group of mice.Flow cytometry was used to detect the effect of mollugin on Hep G2 cell cycle.DCFH-DA staining solution was used to measure the effect of mollugin on Hep G2 cells.DCFH-DA was used to detect small ROS content in mouse H22 tumors.MTT assay was used to detect the inhibitory effect of mollugin on Hep G2 cells treated with ROS inhibitors.Western blot was used to detect cyclin-related proteins and DNA in Hep G2 cells and mouse H22 tissues by western blot Expression of damage marker proteins.ROS inhibitors reversed the expression of mollugin on Hep G2 cells by immunoblotting.RESULTS: The results of MTT assay showed that mollugin significantly inhibited human liver cancer Hep G2 cells in a time-and dose-dependent manner.With the increase of the concentration of the drug,the time of the drug administration was prolonged,and the inhibitory effect gradually became stronger.The results of the plate-cloning assay showed that mollugin reduced the number of clones in a dose-dependent manner.In the overall animal experiment,compared with the model group and the positive control group,the mice in the mollugin administration group had better survival status.The weight of the administration group was close to that of the model group,and the weight of the positive control group was slightly reduced.There was no statistically significant difference in body weight between the model group and the positive control group.High-dose mollugin and adriamycin significantly inhibited tumor growth and significantly reduced the tumor growth trend in mice(P <0.05).High-dose mollugin And adriamycin significantly reduced the final tumor weight(P <0.01,P <0.001).H & E staining results showed that the tumor tissue of the positive control group had obvious necrosis.The tumor cell density of the administration group was reduced,but no obvious necrosis occurred.Liver coefficient results showed that compared with the model group,the liver coefficient of the positive control group was significantly increased(P <0.05),while the mollugin group had a tendency to decrease the organ coefficients without significant differences.Mollugin increased the S-phase ratio of Hep G2 cells in a dose-dependent manner.Mollugin increased the ROS content in Hep G2 cells and mouse H22 tumor tissues in a dose-dependent manner.ROS inhibitors significantly reduced the effect of Mollugin on Hep G2.Inhibitory effect of cells.Cymbalin in dose-dependently reduced Cyclin A2 and CDK2 expression in Hep G2 cells and mouse H22 tumor tissues.Cymbalin in dosedependently increased Hep G2 cells and mouse H22 tumor tissues Expression of DNA damage marker protein p-H2 A.X.ROS inhibitors reversed the change in expression level of related proteins in Hep G2 cell-related proteins.CONCLUSION: Mollugin has shown anti-hepatocellular activity in vivo and in vitro experiments.Its anti-hepatocellular effect may be related to the induction of oxidative stress in cells,resulting in DNA damage and cycle arrest. |
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