Roth J, Goebeler M, van den Bos C, Sorg C
Institute of Experimental Dermatology, University of Münster, Germany.
Biochem Biophys Res Commun. 1993 Mar 15;191(2):565-70. doi: 10.1006/bbrc.1993.1255.
MRP8 and MRP14 are two calcium-binding proteins of the S-100 family which are expressed during distinct stages of monocytic maturation. To further investigate their regulation the human leukemic cell line HL-60 which can be induced to differentiate to monocytes/macrophages by 12-O-tetradecanoylphorbol 13-acetate (TPA), 1,25-(OH)2 Vitamin D3 (VD3), tumor necrosis factor-alpha (TNF-alpha) and interferon-g (IFN-g) were analyzed for expression of MRP8/MRP14. Employing Northern blotting, a sandwich enzyme-linked immunosorbent assay and immunocytochemical analysis we determined MRP8/MRP14 mRNA and protein levels, which were found to be elevated after VD3 and reduced after TPA treatment. TNF-alpha and IFN-g did not affect MRP8/MRP14 levels. Western blot analysis revealed that formation of MRP8/MRP14 to biologically active complexes which has previously been shown to be a calcium-mediated process is not dependent on the differentiation stages of HL-60 cells. Restriction of MRP8/MRP14 expression to only distinct pathways of monocytic differentiation in HL-60 cells may thus reflect different functional phenotypes of monocytes/macrophages in vivo.
MRP8和MRP14是S-100家族的两种钙结合蛋白,在单核细胞成熟的不同阶段表达。为了进一步研究它们的调控机制,我们分析了人白血病细胞系HL-60中MRP8/MRP14的表达情况。HL-60细胞可被12-O-十四酰佛波醇-13-乙酸酯(TPA)、1,25-(OH)2维生素D3(VD3)、肿瘤坏死因子-α(TNF-α)和干扰素-γ(IFN-γ)诱导分化为单核细胞/巨噬细胞。通过Northern印迹法、夹心酶联免疫吸附测定法和免疫细胞化学分析,我们测定了MRP8/MRP14的mRNA和蛋白水平,发现VD3处理后其水平升高,TPA处理后降低。TNF-α和IFN-γ不影响MRP8/MRP14的水平。蛋白质印迹分析表明,MRP8/MRP14形成具有生物活性的复合物(此前已证明这是一个钙介导的过程)并不依赖于HL-60细胞的分化阶段。因此,HL-60细胞中MRP8/MRP14的表达仅局限于单核细胞分化的不同途径,这可能反映了体内单核细胞/巨噬细胞的不同功能表型。