三氧化二砷诱导视网膜母细胞瘤细胞凋亡的实验研究

Arsenic trioxide induced apoptosis in retinoblastoma cells in vitro and its possible mechanism

目的:探讨三氧化二砷体外诱导视网膜母细胞瘤细胞凋亡的作用及其可能的机制。方法:梯度浓度三氧化二砷处理视网膜母细胞瘤细胞系HXO-RB44,MTT法观察增殖的变化,AO/EB染色法、Annexin V+PI法流式细胞仪检测细胞凋亡。活性比色法检测caspase-3活性变化,Western免疫印迹法测定bcl-2和bax蛋白的表达。结果:三氧化二砷处理后的HXO-RB44细胞增殖抑制,抑制率呈剂量时间依赖性;其细胞凋亡率显著增加;caspase-3活性增加,bcl-2/bax的比值下调。结论:三氧化二砷在体外可通过诱导HXO-RB44细胞凋亡达到抑制其增殖的作用;其诱导凋亡的作用可能与激活caspase-3和下调bcl-2/bax比值有关。

Objective To explore the effect of arsenic trioxide on the apoptosis of retinoblastoma cell line HXO-RB44 and the possible mechanism.Methods The effect of arsenic trioxide on the proliferation of retinoblastoma cell line HXO-RB44 was observed by MTT colorimetric assay;the apoptosis of the HXO-RB44 was examined by AO/EB staining and flow cytometry analysis（Annexin V＋ PI staining）;caspase-3 activity and bcl-2/bax expression in the HXO-RB44 were detected by cpp32 colorimetric assay kit and Western blot.Results Arsenic trioxide inhibited the proliferation of HXO-RB44 cell in dose and duration-dependent manner in vitro;arsenic trioxide significantly increased the apoptosis in HXO-RB44 cells.The activation of caspase-3 was increased,and the rate of bcl-2/bax was down-regulated in the HXO-RB44 cells processed with arsenic trioxide.Conclusion Arsenic trioxide can inhibit the proliferation of retinoblastoma cell HXO-RB44 in vitro by apoptosis induction.The apoptosis induction is possibly related to the caspase-3 activation and bcl-2/bax down-regulation.