Radiosensitizing Effect Of4-ANI In HepG2Hepatocellular Carcinoma Cells In Vitro

Objective To study the radiosensitization of4-ANI on HepG2hepatocellularcarcinoma c ell lines in vitro and explore the possible mechanism of cell cycle. Methods Cultured HepG2cell lines in vitro using DMEM, using CCK-8kit for detecting proliferation of HepG2cell line by different concentrations of4-ANI (0,2.5,5,7.5,10,20,30μM) after12h,24h,48h. According to the cell survival rate curve, selecting low cytotoxic drug concentration for subsequent sensitization test. By the cell clone formation assay, re spectively to observe the radiosensitizing effect of10ummol/L,20ummol/Lconcentration of4-ANI on HepG2cell lines.Through the single hit multi target model and the Linear Quadratic Model fitting curve to calculate the related biological parameters. By the B D FACSCALIB detected the changes of HepG2cell cycle distribution after different dos es ofradiation, and the influence of different concentrations of4-ANI on cell cycle of HepG2cells at different time after irradiation. Results In vitro experiments showed tha t theexperimental concentration range of4-ANI had no obvious toxicity on HepG2cells.24hafter drug, cells did not appear obvious inhibition, compared with control group,20uM group and30uM group even promotes cell proliferation;48h after drug, the surviva1curve of the overall downward trend, but shall not be less than80%. And mean sur vival rates of different concentration groups at12,24,48h three time points were statis tically significant,.P<0.001. The clone formation experiment confirmed that4-ANI has obvious radiosensitizing effect on the HepG2cells, and the radiosensitizing effect of4-ANI was positively correlated with Drug concentration. The SERD37of lOμM and20μM treatment groups are1.13and1.30respectively, and SERD10are respectively1.42,1.88.BD FCM testing results showed that radiation can significantly induced HepG2cells arrest in G2/M phase, and the block level and the radiation dose was positive correlatio n trend.4-ANI can decrease the G2arrest of HepG2cells afterradiation, and the ability of4-ANI eliminating G2arrest was positively correlated with drug concentration.At the same time, FCM detected that the percentageof G2phase and S phase was increased on HepG2cells in the simple drug group.Effect of4-ANI on cell cycle of HepG2cell lines after irradiation had no time continuous correlation, only the24h showed significa nt effect of removal of G2phase arrest. The above results by single factor analysis of variance with Holm-Sidakmethod test to analyze the differences between groups werestat istically significant, P<0.05. Conclusion4-ANI medication alone did not produce obvio us toxicity on HepG2cells, and in vitro experiments confirmed that4-ANI has radiosens itizing effect on HepG2cell line, and the sensitization effect was positively correlated with the concentration of4-ANI. FCM detected that the sensitization mechanism was related to the elimination of G2phase arrest on HepG2cells induced by radiation. The se results indicate that PARP inhibitor4-ANI has prospects as radiosensitizing drugs for hepatocellular carcinoma, and provide experimental foundation and theoretical basis for the further study of4-ANI.