As2O3对NB4细胞增殖及凋亡的影响
Effect of arsenic trioxide on apoptosis and cell cycle of NB4 cell
-
摘要:目的 研究不同浓度的三氧化二砷(As2O3)对急性粒细胞白血病细胞株NB4(APLM3)凋亡及细胞周期的影响。方法 采用四甲基偶氮噻唑蓝(MTT)法测As2O3对NB4的细胞毒活性, 流式细胞仪Annexin V FITC-PI检测细胞凋亡率, 溴化丙碇(PI)染色法检测细胞周期。结果 1~8μmol/L As2O3能显著抑制NB4细胞增殖, 且这些变化呈明显的时间和浓度依赖关系(时间-剂量效应)。2~8μmol/L As2O3能显著诱导NB4细胞凋亡, 处理时间为12 h, 其早期凋亡率和总凋亡率均与剂量呈线性正相关; 处理时间为24 h, 其早期凋亡率、中晚期凋亡率和总凋亡率与剂量呈线性正相关; As2O3处理时间为36 h和48 h NB4细胞以中晚期调亡为主。流式细胞检测表明, 不同浓度的As2O3均使NB4细胞周期阻滞在G2/M期。结论 As2O3能抑制人白血病NB4细胞增殖, 其作用机制可能是通过诱导细胞凋亡和抑制细胞增殖使细胞受阻于G2/M期。其诱导凋亡作用具有时间和浓度依赖关系。Abstract:Objective To investigate the changes of apoptosis and cell cycle inducecd by arsenic trioxide(As2O3)inleukemia cell line NB4 cell in vitro.Methods NB4 cells were treated with As2O3 in different co ncentrations fo 12, 24 and 36h.The cytotoxity of NB4 cell were detectexd by(MTT)assay.The apoptotic cell was determined by Annexin VFITC-PI double staining method and measured by flow cytometry(FCM).Cell cycle was examined by PI fluoresence.Results 1~8 Lmol/L As2O3 could induce NB4 cell in a time-and-dose dependent manner in vitro.2~8 Lmol/L As2O3 could induce NB4 cell obvious apoptosis.The apoptosis rate of NB4 cells treated by As2O3 for 12 and 24 h sho wed a line relationship with the concention of As2O3, and for 36 and 48 h the NB4 cell mainly showed later apopotsis.Flow cytometry analysis showed that the number of G2/M phase cells increased gradually with raised concentrations treatment of As2O3.Conclusion As2O3 may inhibit the proliferation of NB4 cell.The mechanism is probably through blocking the cell cycle of G2/M phase and induced NB4 apoptosis in a time-and-dose dependent manner.
下载: