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目的研究脱氧佛波醇乙酸酯(12-deoxyphorbol 13-acetate,Prostratin,DPA)对人急性髓系白血病(AML)细胞株HL-60、NB4和U937细胞分化的影响,并证明其通过激活蛋白激酶C/胞外信号调节激酶(PKC/ERK)通路诱导细胞分化。方法 1μmol/L DPA作用HL-60细胞3 d后观察细胞形态的改变;不同剂量的DPA作用HL-60、NB4、U937细胞24 h后,流式细胞仪检测细胞分化表面标志变化;Western印迹检测ERK蛋白磷酸化的改变;流式细胞仪检测丝裂原活化蛋白激酶激酶(MEK)抑制剂和PKC抑制剂对细胞分化表面标志的影响;Western印迹检测MEK抑制剂和PKC抑制剂对ERK蛋白磷酸化的影响。结果 DPA可诱导HL-60细胞出现白血病细胞分化的形态。DPA剂量依赖性地诱导AML细胞CD11b表达。DPA在诱导分化剂量范围内可剂量依赖性地引起ERK磷酸化。MEK化学小分子抑制剂U0126可完全抑制DPA引起的ERK磷酸化和CD11b表达;PKC非选择性抑制剂GFX可完全抑制DPA引起的ERK磷酸化和CD11b表达。结论 DPA其通过激活PKC/ERK通路能够明显诱导白血病细胞分化。
Objective To investigate the effect of deoxyphorbol 13-acetate (Prostratin, DPA) on the differentiation of human acute myeloid leukemia (AML) cell lines HL-60, NB4 and U937 cells and to prove its effect on the differentiation of human acute myeloid leukemia The kinase C / extracellular signal-regulated kinase (PKC / ERK) pathway induces cell differentiation. Methods HL-60 cells were treated with 1 μmol / L DPA for 3 days. The morphological changes of HL-60 cells were observed after treated with different doses of DPA for 24 h. Flow cytometry was used to detect the changes of cell surface markers. Western blotting ERK protein phosphorylation changes; flow cytometry mitogen-activated protein kinase kinase (MEK) inhibitor and PKC inhibitor on cell differentiation surface markers; Western blot MEK inhibitor and PKC inhibitor of ERK protein phosphorylation The impact of change. Results DPA induced HL-60 cells to differentiate into leukemia cells. DPA induced CD11b expression in AML cells dose-dependently. DPA induced ERK phosphorylation in a dose-dependent manner in the range of induced differentiation doses. MEK chemical small molecule inhibitor U0126 can completely inhibit the DPA-induced ERK phosphorylation and CD11b expression; PKC non-selective inhibitor GFX can completely inhibit the DPA-induced ERK phosphorylation and CD11b expression. Conclusion DPA can obviously induce leukemia cell differentiation through activation of PKC / ERK pathway.