Berg Tobias, Fliegauf Manfred, Burger Jan, Staege Martin S, Liu Shaohua, Martinez Natalia, Heidenreich Olaf, Burdach Stefan, Haferlach Torsten, Werner Milton H, Lübbert Michael
Department of Medicine, Division Hematology/Oncology, University of Freiburg Medical Center, Hugstetter Str. 55, D-79106 Freiburg, Germany.
Haematologica. 2008 Nov;93(11):1728-33. doi: 10.3324/haematol.13044. Epub 2008 Sep 11.
An inducible model for conditional expression of AML1-ETO in myeloid U-937 cells was generated previously to determine cellular effects of AML1-ETO and to identify target genes. Induction of AML1-ETO expression in U-937 resulted in reduced cell growth, G1 arrest and apoptosis. Microarray analysis showed more genes up-regulated than down-regulated (180 vs. 69). Clustering of AML1-ETO-positive and -negative cell lines was possible based on these differentially expressed genes. p21/WAF/Cip1 (CDKN1A) was up-regulated 4.6-fold upon induction of AML1-ETO which was confirmed in additional experiments. Knock-down of AML1-ETO by siRNA could reduce p21/WAF/Cip1 expression in Kasumi-1 cells. mRNA expression analysis of p21/WAF/Cip1 in a large cohort of acute myeloid leukemia patients demonstrated a significantly higher expression in AML1-ETO-positive leukemia. The increased expression of p21/WAF/Cip1 in primary leukemic blasts suggests that elevated p21/WAF/Cip1 levels may contribute to specific features observed in AML1-ETO positive leukemia.
先前构建了一种可诱导模型，用于在髓系U-937细胞中条件性表达AML1-ETO，以确定AML1-ETO的细胞效应并鉴定靶基因。在U-937细胞中诱导AML1-ETO表达导致细胞生长减少、G1期阻滞和凋亡。微阵列分析显示上调的基因比下调的基因更多（180个对69个）。基于这些差异表达基因，可对AML1-ETO阳性和阴性细胞系进行聚类。诱导AML1-ETO后，p21/WAF/Cip1（CDKN1A）上调了4.6倍，这在其他实验中得到了证实。用小干扰RNA敲低AML1-ETO可降低Kasumi-1细胞中p21/WAF/Cip1的表达。对一大群急性髓系白血病患者的p21/WAF/Cip1进行mRNA表达分析显示，在AML1-ETO阳性白血病中其表达显著更高。原发性白血病母细胞中p21/WAF/Cip1表达增加表明，p21/WAF/Cip1水平升高可能导致AML1-ETO阳性白血病中观察到的特定特征。
