摘要
目的:利用Mn(Ⅲ)TBAP能特异性清除细胞内活性氧(reactive oxygen species,ROS),观察不同浓度三氧化二砷(As_2O_3)单独和联合Mn(Ⅲ)TBAP作用于人肝癌HepG_2细胞后细胞内活性氧(ROS)的变化和对凋亡的影响,探讨活性氧(ROS)在三氧化二砷(As_2O_3)诱导肝癌细胞株HepG_2凋亡中的作用。方法:实验分3组:对照组,不同浓度三氧化二砷(As_2O_3)组,三氧化二砷(As_2O_3)+Mn(Ⅲ)TBAP组。用四甲基偶氮唑盐(MTT)比色法测定细胞生长增殖活性;激光共聚焦显微镜荧光染色法检测细胞内ROS的变化;AnnexinV/PI双染色法流式细胞仪检测细胞凋亡。结果:2、4、8μmol/L三氧化二砷(As_2O_3)处理HepG_2细胞后各组细胞增殖抑制率高于对照组(p<0.05),细胞增殖抑制率随As_2O_3浓度的增加及作用时间的延长而升高(p<0.05);HepG_2细胞内ROS的水平随As_2O_3浓度的增加而升高(p<0.05),As_2O_3处理组HepG_2细胞内ROS水平高于空白对照组(p<0.05),As_2O_3+Mn(Ⅲ)TBAP组HepG_2细胞内ROS的水平低于As_2O_3组(p<0.05),但高于对照组(p<0.05)。细胞早期凋亡率随As_2O_3浓度的增加及作用时间的延长而升高(p<0.05),且均高于Mn(Ⅲ)TBAP+As_2O_3组和对照组,Mn(Ⅲ)TBAP+As_2O_3组高于对照组。结论:三氧化二砷作用于HepG_2细胞促进了细胞内ROS的产生,ROS水平的提高增加了三氧化二砷促HepG_2细胞凋亡的敏感性。
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 ceils. The experiment were divided into three groups： the control group, different concentration of arsentic trioxide group and arsentic trioxide combinated Mn（ IU ） 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 （ p 〈 0.05 ） when the concentration of arsentic trioxide is higher than 1 μmol/L and Cell growth inhibitory rate was increased following increasing the concentration of arsentic trioxide and extending the action time （ p 〈 0.05 ）. The level of ROS was markedly inceased and much higher than control group （p 〈 0.05 ）when different concentration （2,4 and 8μmol/L） of arsenic trioxide were used to treat the HepG2 cells. Forepart apoptotic rate were increased（ p 〈 0. 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 HepG2 cells and the level of ROS improved the sensibility of rsentic trioxide inducing HepG2to apoptosis.
出处
《农垦医学》
2009年第5期385-389,共5页
Journal of Nongken Medicine
关键词
三氧化二砷
活性氧(ROS)
人肝癌细胞株HEPG2
凋亡
reactive oxygen species
arsenic trioxide
human hepatocellular carcinoma cell line HepG2
Apoptosisdifferent display
