Stage-specific expression of P2Y receptors, ecto-apyrase, and ecto-5'-nucleotidase in myeloid leukocytes
Stage-specific expression of P2Y receptors, ecto-apyrase, and ecto-5'-nucleotidase in myeloid leukocytes
复制标题
DOI：
10.1152/ajpcell.1997.273.3.c973
复制
发表时间：
1997-09-01
影响因子：
5.5
通讯作者：
Dubyak, GR
中科院分区：
文献类型：
--
作者：
Clifford, EE;Martin, KA;Dubyak, GR
P2嘌呤能受体亚型在白细胞中的表达随谱系和发育阶段而变化。鉴于最近的鉴定和克隆的至少七个不同的G蛋白偶联ATP受体亚型（P2 Y家族），我们研究了P2 Y受体亚型在髓系细胞分化过程中的表达。我们观察到KG-1成髓细胞表达P2 Y（1）而不表达P2 Y（2）受体（以前称为P-2U受体），而后来的髓系祖细胞，包括HL-60早幼粒细胞和THP-1单核细胞，表达P2 Y（2）而不表达P2 Y（1）受体。在KG-1细胞中，P2 Y（1）受体对Ca 2+动员的显著激活作用仅在与外源性ATP酶马铃薯腺苷三磷酸双磷酸酶预孵育后观察到。这表明KG-1细胞中P2 Y（1）受体通过自分泌机制脱敏，该机制可能涉及内源性核苷酸的释放增强和/或细胞表面外核苷酸酶的表达降低。我们比较了KG-1成髓细胞和HL-60早幼粒细胞中胞外腺苷三磷酸双磷酸酶的活性和表达水平。细胞外ATP被HL-60细胞迅速代谢，而KG-1细胞则不能。逆转录-聚合酶链反应分析表明，CD 39（分化簇），一个确定的外腺苷三磷酸双磷酸酶，mRNA存在于HL-60，但KG-1细胞。外三磷酸腺苷双磷酸酶活性随着用佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）或二丁酰腺苷3 '，5'-环一磷酸（DBcAMP）分化髓系祖细胞而适度增加。用PMA而不是DBcAMP诱导HL-60细胞分化，强烈诱导外-5 '-核苷酸酶活性和CD 73 mRNA表达。这些观察结果表明，信号转导细胞外ATP在髓系白细胞中可以调节P2 Y受体亚型和多种外核苷酸酶的表达的发育程序性变化。
The expression of P2 purinergic receptor subtypes in leukocytes varies with both lineage and developmental stage. Given the recent identification and cloning of at least seven distinct G protein-coupled ATP receptor subtypes (P2Y family), we investigated P2Y receptor subtype expression during myeloid cell differentiation. We observed that KG-1 myeloblasts express P2Y(1) but not P2Y(2) receptors (previously termed P-2U receptors), whereas later myeloid progenitors, including HL-60 promyelocytes and THP-1 monocytes, expressed P2Y(2) but not P2Y(1) receptors. In KG-1 cells, significant activation of Ca2+ mobilization by P2Y(1) receptors was only observed after preincubation with potato apyrase, an exogenous ATPase. This indicated that P2Y(1) receptors are desensitized in KG-1 cells by autocrine mechanisms that may involve enhanced release of endogenous nucleotides and/or decreased expression of cell-surface ectonucleotidases. We compared the levels of ecto-apyrase activity and expression in KG-1 myeloblasts and HL-60 promyelocytes. Extracellular ATP was rapidly metabolized by HL-60 but not by KG-1 cells. Reverse transcription-polymerase chain reaction analysis indicated that mRNA for CD39 (cluster of differentiation), an identified ecto-apyrase, was present in HL-60 but not KG-1 cells. Ecto-apyrase activity was modestly increased with differentiation of myeloid progenitors with either phorbol 12-myristate 13-acetate (PMA) or dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP). Differentiation of HL-60 cells with PMA, but not DBcAMP, strongly induced ecto-5'-nucleotidase activity and CD73 mRNA expression. These observations indicate that signal transduction by extracellular ATP in myeloid leukocytes can be regulated by developmentally programmed changes in the expression of P2Y receptor subtypes and multiple ectonucleotidases.