二硫化二砷诱导LY8细胞株凋亡的机制探讨

Exploration on the mechanism of arsenic disulfide-induced LY8 cells apoptosis

目的探讨二硫化二砷诱导弥漫大B细胞淋巴瘤细胞株LY8凋亡的机制。方法将不同浓度的二硫化二砷与弥漫大B细胞淋巴瘤细胞株LY8共培养。采用CCK-8方法测定细胞活性;AnnexinⅤ-PI方法测定细胞凋亡率,采用Western blot技术在蛋白水平测定凋亡相关因子BCL-2、Bax和caspase-3的表达。结果随着二硫化二砷浓度的增高,细胞活性越低,细胞凋亡率越高;随着二硫化二砷作用时间的延长,细胞活性越低,细胞凋亡率也越高。其差异均有统计学意义。Western blot结果显示:与对照组比较,二硫化二砷下调caspase-3蛋白的表达,Bax/BCL-2的比例升高并伴随着Bax断裂。结论二硫化二砷以时间和剂量依赖的方式抑制LY8细胞生长和诱导凋亡,诱导凋亡的机制可能与线粒体通路相关,并伴随着Bax断裂。

Objective To investigate the mechanism of arsenic disulfide （As2S2） on the apoptosis of LY8 human diffuse large B cell lymphoma （DLBCL） cells. Methods LY8 cells were treated with various concentrations of As2S2 for different time periods. Cell viability was evaluated by the CCK-8 assay;Cell apoptosis was detected by AnnexinⅤ-PI analysis. The expression levels of Bax, Bcl-2 and caspase-3 were examined by western blotting. Results We found that the apoptotic rates of LY8 cells were significantly enhanced and the viability of LY8 cells was notably reduced following treatment with As2S2 for 24 h, 48 h and 72 h. Along with increasing As2S2 concentration, the apoptotic rates of LY8 cells were increased and the viability of LY8 cells was decreased. The results were both statistically significant .Western blot analysis revealed that As2S2 increased the Bax/BCL-2 protein ratio in contrast to decreased caspase-3 expression in LY8 cells. Our findings also demonstrated that Bax was proteolytically cleaved in LY8 cells. Conclusions As2S2 inhibited proliferation and induced apoptosis of LY8 cells in a time-and-dose-dependent manner. The effect was partly owing to the induction of mitochondria-dependent apoptosis involving Bax cleavage.