Radiosensitization Effect Of Astragaloside IV On Hepatocellular Carcinoma HepG2 Cells

Objective:To study the effect and mechanism of radiosensitization of astragaloside IV on liver cancer HepG2 cells and its inhibitory effect on liver cancer HepG2 cells,and to provide basis for the development of new tumor radiotherapy sensitizing drugs.Methods:Selection of human liver cancer HepG2 cells,and the cells were treated with astragaloside IV.The experiment was divided into control group,irradiation group,astragaloside IV group,astragaloside IV+irradiation group.The MTT test was used to detect the toxic effects of different concentrations of astragaloside IV on human liver cancer HepG2 cells at different time points,to select the appropriate concentration of astragaloside IV.MTT assay was used to detect changes in cell survival rate at 24,48,and 72 hours after 0,2,4,6,and 8 Gy¹³⁷Cs-γradiation,and to select the appropriate radiation dose.The effect of astragaloside IV on cell viability after irradiation was measured by MTT method;Clone formation experiment to observe the effect of astragaloside IV on the radiosensitivity of HepG2 cells after irradiation;Growth curve to observe the effect of astragaloside on the proliferation of HepG2 cells;Cell scratch test to observe the effect of astragaloside IV on the migration ability of HepG2 cells;Flow cytometry to detect the effect of astragaloside IV on the apoptosis rate and cell cycle of HepG2 cells after irradiation;Western Blot detected the expression of p53,caspase-3,γ-H2AX,Chk2,DNA-PKcs,PI3K,AKT,ATM protein in each group;Micronucleus test detects the damage of cell DNA.Results:1.Compared with the control group,the cell survival rate increased with the increase of astragaloside IV concentration,the longer the action time,the stronger the inhibitory effect on cells.After 48h and72h,compared with the 0μg/ml concentration group,the 80μg/ml concentration group had differences（P<0.05）;Compared with the control group,with the increase of the exposure dose,the time after irradiation increases,the cell survival rate gradually decreases（P<0.05）;Compared with 0μg/ml,the cell survival rate of the irradiated group and astragaloside IV+irradiated group gradually decreased with the increase of drug concentration（P<0.05）,Compared with the irradiation group,the effect of astragaloside IV+irradiation group was stronger（P<0.05）.2.Compared with 0μg/ml,different concentrations of astragaloside IV showed an inhibitory effect on cell growth（P<0.05）;The irradiation group and astragaloside IV+irradiation group can also inhibit the growth of cells（P<0.05）;Compared with the irradiated group,the astragaloside IV+irradiated group had stronger ability to inhibit cell growth and proliferation（P<0.05）.3.Compared with the control group,as the concentration of astragaloside increased,the number of cell clones gradually decreased,and the cell clone survival score gradually decreased（P<0.05）;Compared with 0μg/ml,the formation of clones in the irradiation group and astragaloside IV+irradiation group gradually decreased,and the survival score gradually decreased（P<0.05）;Compared with the irradiation group,the effect of astragaloside IV+irradiation group was stronger（P<0.05）.4.After simply treating with different concentrations of astragaloside IV,Compared with 0μg/ml,the cell migration ability was significantly inhibited,and as the concentration of astragaloside increased,the inhibitory effect on cell migration ability was stronger;Compared with 0μg/ml,the irradiated group and the astragaloside IV+irradiated group also inhibited the cell migration ability,and the astragaloside IV+irradiated group had a stronger effect on inhibiting the cell migration ability.5.Compared with 0μg/ml,as the concentration of astragaloside increased,the micronucleus rate of astragaloside group,irradiation group,astragaloside IV+irradiation group all increased significantly（P<0.05）;Compared with the irradiated group,the micronucleus rate of astragaloside IV+irradiated group was higher（P<0.05）.6.Compared with the control group,astragaloside IV group,irradiation group,astragaloside IV+irradiation group,the apoptosis rate of cells increased with the increase of astragaloside IV concentration（P<0.05）;Compared with the irradiated group,there was a difference in the apoptosis rate between the drug-only group and the IR+80μg/ml group,and the difference was statistically significant（P<0.05）.7.Compared with 0μg/ml,the cell cycle G2/M phase block of the astragaloside IV group at 20 and 80μg/ml began to increase gradually at4h and reached the maximum value at 8h,In the irradiated group and astragaloside IV+irradiated group,the cell cycle G2/M phase block increased more obviously,and began to increase at 4h,and the peak value reached the maximum value at 12h.8.Compared with the control group,with the increase of astragaloside IV concentration,the intracellular expression ofγH2AX,p53,Chk2,and Caspase3 in the astragaloside IV group,irradiation group,and astragaloside IV+irradiation group gradually increased（P<0.05）;Compared with the irradiation group,the protein expression of astragaloside IV+irradiation group was higher（P<0.05）;Compared with the control group,the protein expression of PI3K,AKT,ATM and DNA-PKcs in the astragaloside IV group decreased gradually with the increase of the concentration of astragaloside（P<0.05）;Compared with the irradiated group,the protein expression of PI3K,AKT,ATM,and DNA-PKcs in the astragaloside IV+irradiated group also decreased（P<0.05）.Conclusions:1.Astragaloside IV can inhibit the survival rate of liver cancer cells,which can lead to reduced viability;proliferation and growth ability of liver cancer cells;can reduce the migration ability of liver cancer cells,And the effect of astragaloside IV+irradiation group is stronger.2.Astragaloside IV can induce the apoptosis of liver cancer cells,block the G2/M phase of the cell cycle,can increase the micronucleus rate of cells,And the effect of astragaloside+irradiation group is stronger.3.Astragaloside IV can inhibit the PI3K/AKT signaling pathway and inhibit DNA damage repair,thereby exerting radiosensitizing effect on liver cancer cells.