目的研究早幼粒细胞白血病(PML)蛋白在急性早幼粒细胞白血病(APL)不同阶段存在形式,探讨全反式维甲酸(ATRA)、砷剂、化疗药物对PML蛋白存在形式的影响。方法收集42例APL的骨髓细胞和白血病NB4、MR2细胞,采用1μmol/L三氧化二砷(As2O3)、1μmol/L四硫化四砷(As4S4)、1μmol/L ATRA、10 mg/L阿糖胞苷、1 mg/L高三尖杉酯碱处理,采用间接免疫荧光法,检测处理前后细胞PML荧光信号变化。结果 APL初发病例87.5%表现为小荧光信号(细胞核内密布满天星般荧光亮点,多于30个/细胞),完全缓解病例91.67%表现为大荧光信号(细胞核内有较大的荧光亮点,少于30个/细胞),复发病例中83.33%表现为小荧光信号;与未加药组相比,ARTA组、As2O3组、As4S4组能使初发患者APL细胞的PML蛋白表达由小荧光信号转变为大荧光信号,阿糖胞苷组、高三尖杉酯碱组则无明显影响;ARTA能使NB4细胞PML蛋白表达由小荧光信号转变为大荧光信号,而不能改变MR2细胞PML蛋白表达的小荧光信号,As2O3和As4S4能使NB4、MR2细胞PML蛋白表达由小荧光信号转变为大荧光信号。结论检测APL细胞PML蛋白存在形式对APL预后判断及合理药物选择具有实际应用价值。
Objective To study the expression patterns of promyelocytic leukemia( PML) protein at different stages of acute promyelocytic leukemia( APL) and investigate the effects that the arsenical,all-trans retinoic acid( ATRA) and chemotherapeutic drugs on PML protein expression patterns. Methods The bone marrow cells of 42 APL patients,NB4 cells and MR2 cells were collected and treated separately with 1 μmol/L arsenic trioxide( As2O3),1 μmol/L tetra-arsenic tetra-sulfide( As4S4),1 μmol/L ATRA,10 mg/L cytosine arabinoside,and 1 mg/L homoharringtonine. The changes of fluorescent signals before and after the treatment were detected by indirect immunofluorescence technique. Results The 87. 5% of initial onset APL cases showed small fluorescent signals( fluorescent signals in the nucleus look like a starry sky,more than30 fluorescent light dots per cell); 91. 67% of complete remission APL cases manifested large fluorescent signals( fluorescent signals in the nucleus are larger,fewer than 30 fluorescent light dots per cell); and 83. 33% of relapse APL cases presented small fluorescent signals. Compared with control group without medication,the PML protein expression of APL cells in initial onset cases had larger fluorescent signals in the ATRA group,As2O3 group and As4S4 group,whereas there were no significant changes in the arabinoside cytosine group and the homoharringtonine group. In addition,ATRA transformed the PML protein expression of NB4 cells from small fluorescent signals into large ones,but did not change small fluorescent signals of the PML protein expression in MR2 cells. Also,As2O3 and As4S4 turned the PML protein expression of NB4 and MR2 cells from small fluorescent signals into large ones. Conclusion Detection of PML protein expression patterns of APL cells has practical application values for evaluating the prognosis of APL and making a rational choice of drugs.