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目的 探讨三氧化二砷 (As2 O3 )对人卵巢癌耐药细胞系 3AO/cDDP细胞生长的影响及其机制。方法 采用四甲基偶氮唑蓝 (MTT)法 ,检测不同浓度As2 O3 作用后 ,3AO/cDDP细胞的生长抑制率 ;采用流式细胞技术检测细胞凋亡率 ,细胞周期变化 ,以及Fas、FasL基因表达的变化。所有结果均与人卵巢癌细胞系 3AO细胞相比较 ;采用细胞骨架染色法观察 3AO/cDDP细胞凋亡细胞形态变化 ,通过吖啶橙染色 ,荧光显微镜观察As2 O3 作用后 3AO细胞的形态变化。结果 As2 O3 能明显抑制3AO/cDDP细胞的增殖 ,抑制作用呈时间和剂量依赖性 (P <0 0 5 ) ;在一定浓度范围内 ,3AO/cDDP细胞凋亡率与As2 O3 的浓度和作用时间呈依赖关系 ,诱导凋亡的最适浓度是 3μmol/L ;As2 O3 低浓度时 ,3AO/cDDP细胞周期S期通过受阻 ,高浓度时诱导S期细胞凋亡 ;As2 O3 作用后 ,两种细胞系Fas基因的表达均呈升调节 ,FasL基因的表达无变化 ;与 3AO细胞相比 ,差异均无显著意义 (P >0 0 5 ) ;As2 O3作用后 3AO及 3AO/cDDP形成典型的凋亡小体。结论 As2 O3 通过Fas/FasL系统 ,诱导S期细胞凋亡 ,有效地抑制人卵巢癌耐药细胞系细胞的生长
Objective To investigate the effect of arsenic trioxide (As 2 O 3) on the growth of human ovarian cancer cell line 3AO / cDDP and its mechanism. Methods MTT assay was used to detect the growth inhibition rate of 3AO / cDDP cells treated with different concentrations of As 2 O 3. Flow cytometry was used to detect the apoptosis rate, cell cycle changes, and Fas, FasL Changes in gene expression. All the results were compared with that of human ovarian cancer cell line 3AO. The morphological changes of 3AO / cDDP cells were observed by cytoskeleton staining. The morphological changes of 3AO cells were observed by acridine orange staining and fluorescence microscope. Results As2 O3 significantly inhibited the proliferation of 3AO / cDDP cells in a time and dose-dependent manner (P <0 05). The apoptosis rate of 3AO / cDDP cells and the concentration and duration of As2O3 in a certain concentration range The optimal concentration of As 2 O 3 was 3μmol / L. When the concentration of As 2 O 3 was low, cell cycle S phase of 3AO / cDDP was blocked and the apoptosis of S phase was induced at high concentration. After As 2 O 3 treatment, The expression of Fas gene was up-regulated and the expression of FasL gene was unchanged. Compared with 3AO cells, there was no significant difference (P> 0.05); 3AO and 3AO / cDDP formed typical apoptosis after As2 O3 Small body. Conclusion As2 O3 can induce apoptosis of S phase cells through Fas / FasL system and effectively inhibit the growth of human ovarian cancer cell line