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目的探讨全反式维甲酸(ATRA)联合丙戊酸钠(VPA)诱导HL-60细胞分化过程中BRD4表达调控的分子机制。方法建立ATRA联合VPA诱导白血病细胞株HL-60的分化模型,瑞氏-吉姆萨(Wright-Gimesa)染色观察HL-60细胞形态,流式细胞术检测细胞表面分化抗原的表达,蛋白质印记从蛋白水平检测HL-60细胞经药物诱导24、48、72h后BRD4表达的变化。结果单独应用ATRA或VPA均能够明显抑制白血病细胞生长,诱导细胞分化,倒置显微镜下可观察到HL-60细胞向成熟粒细胞方向分化,细胞表面的分化抗原表达升高,联合诱导分化模型中细胞表面分化抗原表达升高更明显,两者均有统计学意义,在蛋白水平BRD4的表达逐渐降低,联合诱导分化模型中降低更明显,两者有统计学意义。结论 VPA可以明显的促进ATRA诱导白血病细胞的分化作用,BRD4基因的表达在随着分化程度的加强出现逐渐降低的趋势。
Objective To investigate the molecular mechanisms of BRD4 expression induced by all-trans retinoic acid (ATRA) combined with sodium valproate (VPA) in HL-60 cells. Methods The differentiation model of HL-60 cells induced by ATRA and VPA was established. The morphology of HL-60 cells was observed by Wright-Gimesa staining. The expression of cell surface differentiation antigen was detected by flow cytometry. The level of BRD4 expression in HL-60 cells after drug-induced 24,48,72 h was measured. Results Both ATRA and VPA alone could significantly inhibit leukemia cell growth and induce cell differentiation. HL-60 cells differentiated into mature granulocytes and differentiated antigen on the surface of cells were observed under inverted microscope. The expression of surface differentiation antigen was more obvious, both of which were statistically significant. The expression of BRD4 at protein level gradually decreased and the combination of induced differentiation decreased more obviously, both of which were statistically significant. Conclusions VPA can obviously promote the differentiation of leukemia cells induced by ATRA. The expression of BRD4 gradually decreases with the differentiation degree.