| Objective To investigate the effect of Anemoside B4 on the proliferation, migration, invasion and radiosensitivity of gas.tric cancer cells and its molecular mechanism.Methods The study started and ended from April 2018 to October 2019. GES-1 cells and HGC-27 cells were cultured in vitro for 24 h with different concentrations of AB4 (25, 50, 100 μmol/L). Cell viability was deter.mined by MTT assay and the appropriate concentration of AB4 was screened for subsequent studies. The Transwell assay was used todetect the migration and invasion of HGC-27 cells. Cell clone formation assay was used to examine the effect of AB4 on radiosensitivity of HGC-27 cells. Western blotting was used to detect the effect of AB4 on the expression of SIRT6 protein in HGC-27 cells. After inter. fering with SIRT6 expression in combination with AB4 treatment, the above detection methods were used to detect proliferation, migra.tion, invasion and radiosensitivity of HGC-27 cells. The expression levels of DNA-PKcs, Rad51, DNA repair enzymes Ku80, MMP-2 and MMP-9 were detected by Western blotting.Results Compared with the NC group, the survival rate of HGC-27 cells [(100.01± 9.57)% vs. (86.57±6.58)%, (65.45±8.45)%, (49.58±7.96)%] was significantly decreased after AB4 treatment (P<0.05), the number of migrated cells (98.47±8.79 vs. 43.57±6.53) and the number of invasive cells (88.42±9.32 vs. 45.56±5.13) were significantly decreased (P<0.05), and the expression levels of MMP-2 and MMP-9 were significantly decreased(P<0.05), the expression level of SIRT6 protein (0.42±0.03 vs. 1.03±0.15) was significantly increased (P<0.05). Cell clone formation experiments showed that AB4 could decrease the survival fraction of HGC-27 cells (P<0.05), increase the sensitization ratio, and decrease the expression levels of Rad51, DNA-PKcs and Ku80 (P<0.05). After interference with SIRT6 expression and AB4 treatment, the number of migrated cells (44.25±5.52 vs. 86.47± 11.16) and the number of invasive cells (48.56±6.29 vs. 90.17±12.13) increased significantly (P<0.05), the expression levels of MMP-2 and MMP-9 were increased significantly (P<0.05), and the cell survival fraction increased significantly (P<0.05), the expression levels of Rad51, DNA-PKcs and Ku80 protein were significantly increased (P<0.05).Conclusion Anemoside B4 can inhibit the prolifera.tion, migration and invasion of gastric cancer cells, and increase the radiosensitivity of gastric cancer cells by promoting the expressionof SIRT6. |
