一种新型环磷酸腺苷连接的P2受体在未分化HL-60白血病细胞上的药理学特性

Pharmacological profile of a novel cyclic AMP-linked P2 receptor on undifferentiated HL-60 leukemia cells.

作者信息

Conigrave A D, Lee J Y, van der Weyden L, Jiang L, Ward P, Tasevski V, Luttrell B M, Morris M B

机构信息

Department of Biochemistry, University of Sydney, NSW, Australia.

出版信息

Br J Pharmacol. 1998 Aug;124(7):1580-5. doi: 10.1038/sj.bjp.0701985.

DOI:10.1038/sj.bjp.0701985

PMID:9723974

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1565542/

Abstract

Extracellular ATP (EC50=146+/-57 microM) and various ATP analogues activated cyclic AMP production in undifferentiated HL-60 cells. 2. The order of agonist potency was: ATPgammaS (adenosine 5'-O-[3-thiotriphosphate]) > or = BzATP (2'&3'O-(4-benzoylbenzoyl)-adenosine-5'-triphosphate) > or = dATP > ATP. The following agonists (in order of effectiveness at 1 mM) were all less effective than ATP at concentrations up to 1 mM: beta,gamma methylene ATP > or = 2-methylthioATP > ADP > or = Ap4A (P1, P4-di(adenosine-5') tetraphosphate) > or = Adenosine > UTP. The poor response to UTP indicates that P2Y2 receptors are not responsible for ATP-dependent activation of adenylyl cyclase. 3. Several thiophosphorylated analogs of ATP were more potent activators of cyclic AMP production than ATP. Of these, ATPgammaS (EC50=30.4+/-6.9 microM) was a full agonist. However, adenosine 5'-O-[1-thiotriphosphate] (ATPalphaS; EC50=45+/-15 microM) and adenosine 5'-O-[2-thiodiphosphate] (ADPbetaS; EC50=33.3+/-5.0 microM) were partial agonists. 4. ADPbetaS (IC50=146+/-32 microM) and adenosine 5'-O-thiomonophosphate (AMPS; IC50=343+/-142 microM) inhibited cyclic AMP production by a submaximal concentration of ATP (100 microM). Consistent with its partial agonist activity, ADPbetaS was estimated to maximally suppress ATP-induced cyclic AMP production by about 65%. AMPS has not been previously reported to inhibit P2 receptors. 5. The broad spectrum P2 receptor antagonist, suramin (500 microM), abolished ATP-stimulated cyclic AMP production by HL-60 cells but the adenosine receptor antagonists xanthine amine congener (XAC; 20 microM) and 8-sulpho-phenyltheophylline (8-SPT; 100 microM) were without effect. 6. Extracellular ATP also activated protein kinase A (PK-A) consistent with previous findings that PK-A activation is involved in ATP-induced differentiation of HL-60 cells (Jiang et al., 1997). 7. Taken together, the data indicate the presence of a novel cyclic AMP-linked P2 receptor on undifferentiated HL-60 cells.

摘要

细胞外ATP（EC50 = 146±57微摩尔）和各种ATP类似物可激活未分化的HL - 60细胞中环磷酸腺苷（cAMP）的生成。2. 激动剂效力顺序为：ATPγS（腺苷5'-O-[3-硫代三磷酸]）≥BzATP（2'，3'-O-(4-苯甲酰苯甲酰)-腺苷-5'-三磷酸）≥dATP≥ATP。以下激动剂（在1毫摩尔时的效力顺序）在浓度高达1毫摩尔时均比ATP效力低：β，γ-亚甲基ATP≥2-甲硫基ATP＞ADP≥Ap4A（P1，P4-二(腺苷-5')四磷酸）≥腺苷≥UTP。对UTP反应不佳表明P2Y2受体不负责ATP依赖性的腺苷酸环化酶激活。3. 几种ATP的硫代磷酸化类似物是比ATP更强的cAMP生成激活剂。其中，ATPγS（EC50 = 30.4±6.9微摩尔）是完全激动剂。然而，腺苷5'-O-[1-硫代三磷酸]（ATPαS；EC50 = 45±15微摩尔）和腺苷5'-O-[2-硫代二磷酸]（ADPβS；EC50 = 33.3±5.0微摩尔）是部分激动剂。4. ADPβS（IC50 = 146±32微摩尔）和腺苷5'-O-硫代单磷酸（AMPS；IC50 = 343±142微摩尔）可抑制亚最大浓度的ATP（100微摩尔）诱导的cAMP生成。与其部分激动剂活性一致，估计ADPβS可最大程度抑制ATP诱导的cAMP生成约65%。此前尚未报道AMPS可抑制P2受体。5. 广谱P2受体拮抗剂苏拉明（500微摩尔）可消除HL - 60细胞中ATP刺激的cAMP生成，但腺苷受体拮抗剂黄嘌呤胺类似物（XAC；20微摩尔）和8-磺基苯基茶碱（8-SPT；100微摩尔）无此作用。6. 细胞外ATP还可激活蛋白激酶A（PK - A），这与之前的研究结果一致，即PK - A激活参与了ATP诱导的HL - 60细胞分化（Jiang等人，1997年）。7. 综上所述，数据表明未分化的HL - 60细胞上存在一种新型的与cAMP相关的P2受体。