Retinoblastoma protein and CCAAT/enhancer-binding protein β are required for 1,25-dihydroxyvitamin D3-induced monocytic differentiation of HL60 cells
Retinoblastoma protein and CCAAT/enhancer-binding protein β are required for 1,25-dihydroxyvitamin D3-induced monocytic differentiation of HL60 cells
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DOI：
10.1158/0008-5472.can-03-3029
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发表时间：
2004-01-01
期刊：
影响因子：
11.2
通讯作者：
Studzinski, GP
中科院分区：
文献类型：
--
作者：
Ji, Y;Studzinski, GP
众所周知，维生素D的衍生物（类三角洲）具有诱导多种恶性细胞分化的能力，包括人类白血病细胞，但导致这种结果的信号通路尚不清楚。在这项研究中，我们研究了视网膜母细胞瘤蛋白（pRb）和CCAAT/增强子结合蛋白（C/EBP） 0在1,25-二羟基维生素D-3 （1,25d(3)）诱导的人白血病HL60细胞单核细胞分化中的作用。研究发现，在该系统中，pRb在暴露于诱导剂12小时内上调，其增加的动力学与早期分化标志物CD14和单核细胞特异性酯酶的出现相似。pRb表达的增加伴随着C/EBP，6蛋白的类似增加，这两种蛋白共免疫沉淀，提示形成复合物。与pRb或C/EBPbeta反义的寡核苷酸（但不与C/EBPalpha反义）或含有C/ ebp结合序列的寡核苷酸（“诱饵”）都能抑制1,25 d(3)诱导的分化。分别使用药物抑制剂ZK159222、PD98059或SP600125抑制维生素D受体或丝裂原活化蛋白激酶（MAPK）胞外信号调节激酶和C -jun- nh2末端激酶途径的信号传导，以协调的方式抑制pRb和C/ fbp β的表达和分化。相反，SB202190抑制p38MAPK通路可增强pRb和C/EBPbeta的分化和上调。我们认为1,25D(3)可能通过维生素D受体依赖的方式向HL60细胞的单核细胞分化发出信号，包括激活细胞外信号调节激酶和C - jun - nh2末端激酶MAPK途径，从而上调pRb和C/ fbp β的表达，进而启动分化过程。
Derivatives of vitamin D (deltanoids) are well known to have the ability to induce differentiation of a variety of malignant cells, including human leukemia cells, but the signaling pathways that lead to such an outcome are unclear. In this study we investigated the role of the retinoblastoma protein (pRb) and the CCAAT/enhancer-binding protein (C/EBP) 0 in 1,25-dihydroxyvitamin D-3 (1,25D(3))-induced monocytic differentiation of human leukemia HL60 cells. It was found that in this system, pRb is up-regulated within 12 h of exposure to the inducer, and the kinetics of its increase parallel the appearance of the early markers of differentiation, CD14 and monocyte-specific esterase. The increase in pRb expression was accompanied by a similar increase in C/EBP,6 protein, and these two proteins coimmunoprecipitated, suggesting formation of a complex. Oligonucleotides antisense to pRb or C/EBPbeta (but not to C/EBPalpha) or containing the C/EBP-binding sequence ("decoys"), all inhibited 1,25D(3)-induced differentiation. Inhibition of signaling by vitamin D receptor or by mitogen-activated protein kinase (MAPK) extracellular signal-regulated kinase and c-jun-NH2-terminal kinase pathways using pharmacological inhibitors ZK159222, PD98059, or SP600125, respectively, inhibited pRb and C/FBPbeta expression and differentiation in a coordinate manner. In contrast, inhibition of the p38MAPK pathway by SB202190 potentiated differentiation and the up-regulation of pRb and C/EBPbeta. We suggest that 1,25D(3) may signal monocytic differentiation of HL60 cells in a vitamin D receptor-dependent manner that includes activation of extracellular signal-regulated kinase and c-Jun-NH2-terminal kinase MAPK pathways, which then up-regulate pRb and C/FBPbeta expression and in turn initiate the differentiation process.