HL-60细胞分化与凋亡关系的初步研究

目的探讨HL-60细胞分化与凋亡的初步关系,指导白血病的联合化疗.方法NBT还原试验检测细胞分化、流式细胞术(FCM)检测细胞凋亡及bcl-2、c-myc蛋白的表达,观察分别经ATRA、TPA诱导分化的HL-60细胞对VP-16的凋亡反应.结果与对照组相比,经ATRA、TPA分化的HL-60细胞bcl-2、c-myc蛋白表达都明显降低(P＜0.05),其中以TPA下调c-myc蛋白的幅度最大(76%);对VP-16的诱导反应,经TPA分化的HL-60细胞凋亡率明显减少(P＜0.05),而经ATRA分化的细胞凋亡率则无明显变化(P＞0.05).结论HL-60细胞对VP-16的凋亡诱导反应与bcl-2、c-myc蛋白的下调表达程度有关;分化诱导剂ATRA与低剂量的VP-16联合应用可望改善AML的化疗效果,并且以后者先用效果更好.

STUDY ON THE RELATIONSHIP BETWEEN DIFFERENTIATION AND APOPTOSIS OF HL-60 CELLS

Objective:Explore the relationship between differentiation and apoptosis of HL-60 cells,to direct the chemiotherapy of acute leukemia.Methods:Use NBT test to detect cell differentiation,flow cytometic (FCM)to analysis apoptotic cells and the expression of bcl-2 and c-myc protein.So study the apoptosis response induced by etoposide (VP-16)in adherent ATRA and TPA-differentiated HL-60 cells and its probably mechanism.Results:The gene expression of bcl-2 and c-myc was significantly lower in ATRA and TPA-differentiated HL-60 cells than that in control( P<0.05 ),c-myc protein levels down-regulated by TPA was the lowest(76%).Compared with control HL-60 cells,the percent of apoptotic cells induced by VP-16 in adherent TPA-differentiated HL-60 cells was apparently decreased ( P<0.05 ),but such changes can't be found in adderent ATRA-differentiated HL-60 cells ( P>0.05 ).Conclusion:The apoptotic respone induced by VP-16 is different in differentiated HL-60 cells and this perhaps due to the down-regulated levels of bcl-2 and c-myc protein.Differentiated drugs such as ATRA combined with low dose of VP-16 can improve the chemotherapy efficiency of AML,while the latter administrated first is better probably.