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目的研究蟾蜍灵与全反式维甲酸(ATRA)联合对急性早幼粒细胞白血病(APL)原代培养细胞的诱导分化作用。方法12例初诊的APL患者的骨髓经单个核细胞分离后进行原代培养。细胞存活率测定采用锥虫蓝拒染法,细胞凋亡及诱导分化采用形态学分析、四唑氮蓝(NBT)还原试验及细胞分化抗原CD11b的流式细胞仪分解。结果蟾蜍灵5nmol/L与ATRA30nmol/L联合应用明显诱导APL原代培养细胞向成熟粒细胞分化、联合应用较单独应用ATRA时NBT还原率增加了15%～52%及CD11b表达增加了16%～69%,同时分化时间从7d缩短至4d,而且ATRA用量可减少至单用时的30%。结论蟾蜍灵与ATRA联合应用可显著增强ATRA对APL原代培养细胞的诱导分化作用。
Objective To investigate the differentiation of primary cultured acute promyelocytic leukemia (APL) cells induced by bufalin and all-trans retinoic acid (ATRA). Methods The bone marrow of 12 newly diagnosed APL patients was isolated from mononuclear cells and then primary cultured. Cell viability was determined by trypan blue exclusion. Morphological analysis, NBT reduction assay and flow cytometric analysis of CD11b were used to detect apoptosis. Results The combination of Bufalin 5 nmol / L and ATRA 30 nmol / L significantly induced APL primary cultured cells to differentiate into mature granulocytes. Compared with ATRA alone, the addition of Bufalin increased the NBT reduction rate by 15% -52% and the expression of CD11b by 16% 69%, while the differentiation time from 7d to 4d, and the amount of ATRA can be reduced to 30% alone. Conclusion Bufalin and ATRA can significantly enhance the induction of ATRA on APL primary cultured cells.