雄黄诱导维甲酸耐药的急性早幼粒细胞白血病细胞(MR2细胞)凋亡的体外研究

目的 :深入了解雄黄治疗急性早幼粒细胞白血病 (APL)的机理。方法 :以对全反式维甲酸 (ATRA)耐药的APL细胞株MR₂ 细胞为体外模型 ,用不同剂量的雄黄处理细胞一定时间后 ,利用MTT实验观察细胞的存活、增殖状态 ,应用细胞形态学、基因组DNA电泳、流式细胞仪检测细胞凋亡 ,利用流式细胞仪分析细胞周期及细胞膜AnnexinⅤ的表达。结果 :一定浓度范围内雄黄对MR₂ 细胞的生长、增殖有剂量、时间依赖性抑制作用 ;Wright’s及HE染色均可见凋亡细胞 ,透射电镜下可见核染色质边集、核碎裂及凋亡小体 ;基因组DNA电泳出现“梯形”条带 ;流式细胞仪分析出现低于G1期DNA含量的亚二倍体细胞峰 ;经雄黄处理后AnnexinⅤ FITC⁺/PI^-的凋亡细胞逐渐增多。结论 :雄黄可诱导ATRA耐药的APL细胞发生凋亡 ,提示雄黄治疗APL的效应途径可能不同于ATRA。

In vitro Study on Realgar Induced Apoptosis of All-trans Acid Resistant Acute Promeylocytic Leukemia Cell Line (MR2)

OBJECTIVE: To acquire a deep understanding of the possible mechanisms of realgar in the treatment of acute promyelocytic leukemia (APL). METHOD: All-trans retinoic acid (ATRA) resistant APL cell line MR2 was used as in vitro model. The effect of realgar on MR2 cell was observed by watching cell viability, cell growth, and by using Methy thiazolyl tetrazolium (MTT) assay, cell morphology, DNA gel electrophoresis and flow cytometry assay. RESULT: The viability and growth of MR2 cell were inhibited after the treatment, to some extent, in a dose and time dependent manner. After being treated with realgar, MR2 cell presented morphologically some features of apoptotic cells such as intact cell membrane, chromatin condensation and nuclear fragmentation, and apoptotic body could be found by electron microscopy as well. Sub-G1 cells were observed by flow cytometry, as well as Annexin V FITC+/PI-cells. DNA ladder could be found by DNA gel electrophoresis. CONCLUSION: Realgar can induce apoptosis of ATRA resistant APL cell line MR2, Which shows the therapeutic effect of realgar on APL may be different from that of ATRA.