摘要
目的研究三氧化二砷（As2O3）诱导人肝癌细胞系Bel-7402细胞凋亡及与线粒体跨膜电位的关系。方法应用不同浓度的As2O3作用于肝癌细胞后，观察As2O3对肝癌细胞生长状态的影响；通过噻唑蓝（MTT）比色法观察其对Bel-7402细胞增殖的影响；利用共聚焦显微镜及流式细胞术观察经Annexin V-FITC／PI双染后的细胞早期凋亡；通过共聚焦显微镜及流式细胞仪检测线粒体跨膜电位（MMP）的改变情况；并通过底物染色法反映Caspase-3的活性。结果不同浓度的As2O3作用于肝癌细胞后有明显的时间和剂量依赖性；经Annexin V-FITC／PI双染后，可观察到Bel-7402细胞的早期凋亡现象，2μmol／L As2O3作用24h细胞凋亡率为9．89％，作用48h细胞凋亡率为48．53％，而4μmol／L As2O3作用24h细胞凋亡率为18．27％，作用48h细胞凋亡率为67．52％；经As2O3药物作用后，细胞线粒体膜电位水平下降，且与药物作用时间和药物浓度有关（P〈0．05）；同时Caspase-3活性被激活。结论As2O3可明显抑制肝癌细胞Bel-7402的生长，并使细胞线粒体膜电位下降，诱导肝癌细胞凋亡。
Objective To investigate the relationship between Bel-7402 cell apoptosis induced by arsenic trioxide （As2O3） and cell mitochondrial membrane potential. Methods After treatment with As2O3 of different concentrations （0.5-8 μmol/L）, the effect of As2O3 on the growth of liver cancer cell line Bel-7402 was observed. The inhibitory effect of As2O3 on the cell proliferation was investigated with 2-（4,5-dimethyl-thizazol-2-yl）-2,5 diphenyl tetrazolium bromide （MTT） method and its inducibility of cell apoptosis detected by confocal microscopy and flow cytometry after dying with Annexin V-FITC/PI. The mitochondrial membrane potential was determined by confocal microscopy and flow cytometry and the activation of caspase-3 was assayed by substrate color reaction. Results The effect of As2O3 on liver cancer cell.line Bel-7402 was dependent on the time and concentration obviously. The apoptotic cells were detected by confocal microscopy and flow cytometry after dying with Annexin V-FITC/PI. The mitochondrial membrane potential decreased in a dose-time dependent manner and the caspase-3 was activated. Conclusions As2O3 can significantly inhibit the growth of liver cancer cell line Bel-7402, induce its apoptosis and decrease the mitochondrial membrane potential.
出处
《中华肝胆外科杂志》
CAS
CSCD
2007年第10期698-700,共3页
Chinese Journal of Hepatobiliary Surgery
基金
本课题研究受国家自然科学基金（30300339）资助
