Relationship between reactive oxygen species and apoptosis in HepG2 cells induced by Arsenic Trioxide(As_2O_3)

Objective:To investigate the effects of reactive oxygen species on apoptosis of HepG2 induced by arsenic trioxide by use of the Mn(Ⅲ) TBAP can specificly eliminate ROS in cells.Methods:Different concentration(2、4 and 8μmol/L) of arsenic trioxide and Mn(Ⅲ) TBAP were used to treat the HepG2 cells.The experiment were divided into three groups:the control group,different concentration of arsentic trioxide group and arsentic trioxide combinated Mn(Ⅲ) TBAP group.MTT assay was used to examine the cell growth inhibitory rate,Laser sanning confocal microscope(LSCM) was used to examine the variation of reactive oxygen species and AnnexinV/PI double labeling FCM was used to determine cell apoptotic rate.Results:Cell growth inhibitory rate of each group was higher than control group(p0.05) when the concentration of arsentic trioxide is higher than 1μmol/L and Cell growth inhibitory rate was increased following increasing the concentration of arsentic trioxide and extending the action time(p0.05).The level of ROS was markedly inceased and much higher than control group(p0.05) when different concentration(2、4 and 8μmol/L) of arsenic trioxide were used to treat the HepG_2 cells.Forepart apoptotic rate were increased(p0.05) following the raised of the concentration of arsenic trioxide and the extention of action time.fuethermore,both higher than the control group and arsentic trioxide combinated Mn(Ⅲ) TBAP group,the control group was higher than rsentic trioxide combinated Mn(Ⅲ) TBAP group.Conclusion: More reactive oxygen species was produced when rsentic trioxide were used to treat the HepG_2 cells and the level of ROS improved the sensibility of rsentic trioxide inducing HepG_2 to apoptosis.