2-methoxyestradiol-induced apoptosis in U937 cells through generation of reactive oxygen species / 吉林大学学报(医学版)

ABSTRACT

Objective To investigate the effect of 2-methoxyestradiol(2-ME) on U937 myeloid leukemia cell line and its mechanism.Methods The experiment was divided into control group(myeloid leukemia U937 cell in RPMI 1640 culture medium with equal DMSO),2-ME-treated group,NAC-treated group,and 2-ME+NAC-treated group.The cytotoxicity was analyzed by MTT assay.Apoptosis and cellular nitric oxide(NO) were detected by flow cytometry using annexin V and NO sensor dye.Superoxide anion was measured with a fluorescent plate reader by DHE.Results Viabilities of U937 cells treated with 2-ME(0.25,0.50,1.00,and 2.00 ?mol?L-1) for 48 h were gradually reduced to 0.68?0.05,0.28?0.07,0.18?0.07,and 0.11?0.04,respectively.The differences were significant compared with control group(1.00?0.05)(P0.05).2.00 ?mol?L1 2-ME also significantly increased the mean fluorescence of NO sensor dye and superoxide anions in U937 cells compared with control group(P0.05).An markedly increase in apoptotic cells was detected after 2-ME treatment for 3 h(6.78%?1.01% vs 1.59%?0.12%,P