PDF(Pubmed)
Abstract：
Differentiation therapy using all-trans retinoic acid (ATRA) for acute promyelocytic leukemia (APL) is well established. However, because the narrow application and tolerance development of ATRA need to be improved, we searched for another efficient myeloid differentiation inducer. Kinase activation is involved in leukemia biology and differentiation block. To identify novel myeloid differentiation inducers, we used a Kinase Inhibitor Screening Library. Using a nitroblue tetrazolium dye reduction assay and real-time quantitative PCR using NB4 APL cells, we revealed that, PD169316, SB203580, SB202190 (p38 MAPK inhibitor), and triciribine (TCN) (Akt inhibitor) potently increased the expression of CD11b. We focused on TCN because it was reported to be well tolerated by patients with advanced hematological malignancies. Nuclear/cytoplasmic (N/C) ratio was significantly decreased, and myelomonocytic markers (CD11b and CD11c) were potently induced by TCN in both NB4 and acute myeloid leukemia (AML) M2 derived HL-60 cells. Western blot analysis using NB4 cells demonstrated that TCN promoted ERK1/2 phosphorylation, whereas p38 MAPK phosphorylation was not affected, suggesting that activation of the ERK pathway is involved in TCN-induced differentiation. We further examined that whether ATRA may affect phosphorylation of ERK and p38, and found that there was no obvious effect, suggesting that ATRA induced differentiation is different from TCN effect. To reveal the molecular mechanisms involved in TCN-induced differentiation, we performed microarray analysis. Pathway analysis using DAVID software indicated that \"hematopoietic cell lineage\" and \"cytokine-cytokine receptor interaction\" pathways were enriched with high significance. Real-time PCR analysis demonstrated that components of these pathways including IL1β, CD3D, IL5RA, ITGA6, CD44, ITGA2B, CD37, CD9, CSF2RA, and IL3RA, were upregulated by TCN-induced differentiation. Collectively, we identified TCN as a novel myeloid cell differentiation inducer, and trials of TCN for APL and non-APL leukemia are worthy of exploration in the future.
摘要：
使用全反式维甲酸（ATRA）治疗急性早幼粒细胞白血病（APL）的分化治疗已经确立。然而,由于ATRA的狭窄应用和耐受性发展有待改进，我们寻找另一种有效的髓样分化诱导剂.激酶激活参与白血病生物学和分化阻滞。为了鉴定新的髓样分化诱导剂，我们使用了激酶抑制剂筛选库。使用硝基蓝四唑染料还原测定法和使用NB4APL细胞的实时定量PCR，我们透露，PD169316,SB203580,SB202190(p38MAPK抑制剂),和曲西瑞宾（TCN）（Akt抑制剂）有效增加CD11b的表达。我们专注于TCN，因为据报道晚期血液系统恶性肿瘤患者对TCN的耐受性良好。核/细胞质(N/C)比值显著降低，在NB4和急性髓性白血病（AML）M2衍生的HL-60细胞中，TCN均有效诱导了骨髓单核细胞标志物（CD11b和CD11c）。使用NB4细胞的蛋白质印迹分析表明TCN促进ERK1/2磷酸化,而p38MAPK磷酸化不受影响，提示ERK途径的激活与TCN诱导的分化有关。我们进一步研究了ATRA是否可能影响ERK和p38的磷酸化，发现没有明显的影响，表明ATRA诱导的分化不同于TCN效应。为了揭示TCN诱导分化的分子机制，我们进行了微阵列分析.使用DAVID软件的通路分析表明，“造血细胞谱系”和“细胞因子-细胞因子受体相互作用”通路具有很高的意义。实时PCR分析表明，这些途径的成分包括IL1β，CD3D,IL5RA,ITGA6,CD44,ITGA2B,CD37,CD9,CSF2RA,和IL3RA，通过TCN诱导的分化上调。总的来说，我们确定TCN是一种新的骨髓细胞分化诱导剂，TCN治疗APL和非APL白血病的试验值得进一步探索。
