摘要
目的探讨三氧化二砷(As2O3)对人肝癌细胞系HepG2的凋亡诱导作用及其发生机制。方法体外培养HepG2细胞,用不同浓度的As2O3对HepG2细胞进行干预;采用MTT、Annexin V/PI双染及TUNEL法,观察其对HepG2细胞的增殖抑制和凋亡诱导作用;采用流式细胞术定量检测凋亡相关蛋白Bcl-2与Bax表达的变化。结果 As2O3在体外能明显抑制HepG2细胞生长,具有时间剂量依赖关系。Annexin V/PI双染及TUNEL法结果显示:5～20μmol/L的As2O3作用24h均可诱导HepG2细胞凋亡,且呈剂量依赖性,差异均有统计学意义(t分别=-2.96、-4.15、-7.33,P均<0.05);As2O3作用HepG2细胞24h时Bcl-2表达下降,Bax表达上升,Bcl-2/Bax比值降低,呈剂量依赖性,差异均有统计学意义(t分别=3.59、5.65、6.94、7.62、7.92,P均<0.05)。结论一定浓度的As2O3能抑制HepG2细胞增殖,促进其凋亡,其机制可能与调控Bcl-2、Bax表达有关。
Objective To investigate the apoptosis inducing activity of arsenic trioxide in hepatoma cell line HepG2 and its mechanisms.Methods HepG2 cells were treated with arsenic trioxide at 1.25,2.5,5,10,20μmol/L.Cell growth inhibitory rate was detected by MTT colorimetric assay every 24 hours.Apoptosis rate was analyzed by Annexin V/PI fluorescence staining and TUNEL.Changes in expression of Bcl-2 and Bax protein were assayed by flow cytometry.Results MTT tests showed HepG2 cell proliferation was significantly inhibited by arsenic trioxide in time and dosedependent fashion.Marked apoptosis was detected by Annexin V/PI and TUNEL in HepG2 cells after treatment by arsenic trioxide at 5,10,20μmol/L after 24 hours and increased in a dose-dependent manner（t=-2.96,-4.15,-7.33,P 0.05）.The Bcl-2 expression decreased and the Bax expression increased in HepG2 cells induced by arsenic trioxide after 24 hours （t=3.59,5.65,6.94,7.62,7.92,P0.05）.Conclusions Bcl-2 and Bax protein probably correlated with the activation of apoptosis in HepG2 cells induced by arsenic trioxide.
出处
《全科医学临床与教育》
2011年第5期490-493,共4页
Clinical Education of General Practice
