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目的探讨三氧化二砷(As2O3)诱导K562细胞凋亡的发生机制。方法分别以1、2、4、8、16μmol/LAs2O3诱导K562细胞凋亡,MTT法检测不同时间点细胞增殖抑制率,DNA Ladder法检测细胞凋亡,Annexin V/PI染色结合流式细胞术检测细胞凋亡率,Rhodamine123染色流式细胞术检测线粒体膜电位(ΔΨm)变化,ELISA法检测胞浆细胞色素C(CYTC)水平,分光光度法检测Caspase-3活性,流式细胞术检测Bcl-2和Bax表达。结果MTT结果显示As2O3能抑制K562细胞生长,且呈现时间剂量依赖性;4~16μmol/L的As2O3作用24h均可诱导K562细胞凋亡,同时伴有ΔΨm下降,胞浆内CYTC蛋白浓度及Caspase-3活性增高,胞浆Bcl-2/Bax值下降,且均呈剂量依赖性。结论As2O3诱导K562细胞凋亡可能是通过降低ΔΨm,激活线粒体凋亡途径实现的,Bcl-2/Bax值下降可能与其有关。
Objective To investigate the mechanism of apoptosis induced by As2O3 in K562 cells. Methods K562 cells were induced with As2O3 at 1, 2, 4, 8 and 16μmol / L, respectively. MTT assay was used to detect the cell proliferation inhibition at different time points. DNA ladder assay was used to detect apoptosis. Annexin V / PI staining combined with flow cytometry (A|Ìm) were detected by Rhodamine 123 staining, cytoplasmic cytochrome C (CYTC) level by ELISA, Caspase-3 activity by spectrophotometry and Bcl-2 by flow cytometry And Bax expression. Results MTT assay showed that As2O3 could inhibit the growth of K562 cells in a dose-and time-dependent manner. Apoptosis of K562 cells induced by As2O3 at 4 ~ 16μmol / L for 24 h could induce apoptosis of K562 cells with decreased ΔΨm, CYTC protein concentration and Caspase- 3 activity increased, cytoplasmic Bcl-2 / Bax values decreased, and were dose-dependent. Conclusion As2O3 induced K562 cell apoptosis may be through the mitigation of ΔΨm activation of mitochondrial apoptosis pathway, Bcl-2 / Bax value may be related to its decline.