二乙酰二脱水卫矛醇诱导人白血病HL-60细胞凋亡与Bcl-2家族的关系

The relationship between diacetyldianhydrogalactitol-induced apoptosis in HL-60 cells and Bcl-2 family

目的 探讨抗肿瘤药物二乙酰二脱水卫矛醇 (di acetyldianhydrogalactitol,DADAG )诱导肿瘤细胞凋亡机制。方法 透射电镜、DNA梯形条带、流式细胞术及Westernblot法分别检测凋亡和凋亡相关基因Bcl 2家族成员表达的动态变化。结果 透射电镜可见细胞异染色质固缩、边集及凋亡小体形成 ;DNA琼脂糖凝胶电泳呈典型的“DNAlad der” ;流式细胞仪DNA直方图上出现亚二倍体峰。DADAG4 μmol·L-1处理HL 6 0细胞 6h后 ,Bcl 2蛋白水平开始下降 ,Bad蛋白水平明显上调 ,相反 ,Bcl XL 蛋白水平呈时间依赖性地下降。结论 DADAG诱导白血病HL 6 0细胞凋亡 ,与Bcl 2、Bcl XL 蛋白水平下降 。

AIM To investigate the apoptotic activity induced by diacetyldianhydrogalactitol (DADAG) and its mechanism in human leukemia HL 60 cells. METHODS DADAG induced apoptosis in HL 60 cells was observed by electron microscopy, flow cytometry and DNA fragmentation assay. The levels of Bcl 2 family proteins were detected by Western blot. RESULTS HL 60 cells showed typical apoptosis features: nuclear condensation, margination against the nuclear envelope and apoptotic body, typical DNA ladder, subdiploid peak before G 0/G 1 phase was observed by flow cytometry. After treatment with DADAG 4 μmol·L -1 the levels of Bcl 2 protein and Bcl X L protein decreased in a time dependent manner, while the level of Bad protein was increased. CONCLUSION DADAG induced apoptosis correlated with down regulation of Bcl 2 and Bcl X L and up regulation of Bad in human leukemia HL 60 cells.