Biological Characteristics Effect Of PARP-1 Inhibitor On Human Hepatoma Cell LineHepG2

ObjectTo investigate the effect of three different PARP-1 inhibitors on biological characteristics of human hepatoma cell line HepG2 cells,preliminary discussion on the possible mechanism of proliferation and apoptosis of HepG2 cells induced by PARP-1 inhibitors, and provide a new therapeutic target for liver cancer.MethodsTo observe the suppressive effects of different concentrations of three PARP-1 inhibitors (AG014699,BSI-201,AZD-2281) on the proliferation of human hepatoma cell line HepG2 cells were determined by the MTT assay. Selecting two sensitive PARP-1 inhibitors AG014699 and BSI-201, the effect of the two sensitive PARP-1 inhibitors on apoptosis in HepG2 cell was detected by flow cytometry(FCM). The expressions levels of Casepase3,Casepase8,Bax and Bcl-2 were detected by western blotting. The effect of human hepatoma cell line HepG2 cells migration of AGO 14699 and BSI-201 was examined by Transwell assay.ResultsThe results indicated that three different PARP-1 inhibitors(AGO 14699, BSI-201,AZD-2281 Suppressed the growth of HepG2 cells in a time and dose dependent manner, however, the sensitivity of three kinds of PARP-1 inhibitors to HepG2 cells is different. The IC50 is about 20μmol/l,30μmol,400μmol/l treated by them after 48h. Using flow cytometry and Western Blot assay to detect two sensitive PARP-1 inhibitors AG014699 and BSI-201 induced apoptosis of HepG2 cells. With 10μmol/l,30μmol/l, 50μmol/l AGO 14699 and 20μmol/1,40 μmol/1,60μmol/1 BSI-201 induced apoptosis of HepG2 cells, and the apoptosis rate of AGO 14699 and BSI-201 treated HepG2 cells was significantly higher than the control cells at 48 hours,there was significant difference(31% vs 0.01%,t=15.019,P<0.01;24.12% vs 0.03%, t=30.537, P<0.01). We found that AGO 14699 and BSI-201 significantly increases Caspase3, Caspase8 and Bax in HepG2 cells, but decreases bcl-2 protein level in HepG2 cells relative to the control group,and there was a significant difference relative to the control group (P<0.01),figure3A and 3B.ConclusionsThree kinds of PARP-1 inhibitors could significantly inhibit the proliferation of human hepatoma cell line HepG2 cells, but different sensitivity, AGO 14699 and BSI-201 can induce apoptosis of HepG2 cells. Three kinds of PARP-1 inhibitors exhibited different sensitivity on HepG2 cell, with the relative potencies of AG014699> BSI-201> AZD-2281, AG014699 is the most sensitive on HepG2 cells, followed by BSI-201, AZD-2281 is not sensitive. Our study provides a preliminary experimental evidence by using PARP-1 inhibitors as a new target for the treatment of hepatoma. And put forward a new ideas and methods for the clinical treatment of liver cancer.