Ecto-nucleoside triphosphate diphosphohydrolase 2 modulates local ATP-induced calcium signaling in human HaCaT keratinocytes.
Ecto-nucleoside triphosphate diphosphohydrolase 2 modulates local ATP-induced calcium signaling in human HaCaT keratinocytes.
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DOI：
10.1371/journal.pone.0057666
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Lin CH
中科院分区：
文献类型：
--
作者：
Ho CL;Yang CY;Lin WJ;Lin CH
角质形成细胞是人类表皮的主要组成部分。在许多生理和病理生理条件下，角质形成细胞释放三磷酸腺苷 (ATP) 作为自分泌/旁分泌介质，调节细胞增殖、分化和迁移。 ATP 受体已在多种表皮细胞类型中被发现；因此，细胞外 ATP 稳态可能决定其对皮肤健康的长期营养影响。我们研究了人类角质形成细胞在细胞周微环境中表达调节 ATP 浓度以及相应受体激活的表面酶的可能性。我们观察到人角质形成细胞系 HaCaT 释放 ATP 并水解细胞外 ATP。有趣的是，ATP 水解导致二磷酸腺苷 (ADP) 在细胞外空间积聚。 ARL 67156 的药理抑制或内源性外核苷三磷酸二磷酸水解酶 (NTPDase) 亚型 2 的基因沉默可导致 ATP 水解和 ADP 形成减少 25%。使用细胞内钙作为报告基因，我们发现虽然 NTPDase2 水解 ATP 并产生可持续的 ADP 水平，但只有 ATP 通过 P2Y2 受体激活而有助于增加细胞内钙。此外，敲低 NTPDase2 增强了纳摩尔 ATP 诱导的细胞内钙增加，表明 NTPDase2 整体减弱了细胞周微环境中的核苷酸浓度，并局部屏蔽了附近的受体免受细胞外 ATP 的激活。我们的研究结果揭示了人角质形成细胞 NTPDase2 在调节人表皮细胞外环境中的核苷酸信号传导中的重要作用。
Keratinocytes are the major building blocks of the human epidermis. In many physiological and pathophysiological conditions, keratinocytes release adenosine triphosphate (ATP) as an autocrine/paracrine mediator that regulates cell proliferation, differentiation, and migration. ATP receptors have been identified in various epidermal cell types; therefore, extracellular ATP homeostasis likely determines its long-term, trophic effects on skin health. We investigated the possibility that human keratinocytes express surface-located enzymes that modulate ATP concentration, as well as the corresponding receptor activation, in the pericellular microenvironment. We observed that the human keratinocyte cell line HaCaT released ATP and hydrolyzed extracellular ATP. Interestingly, ATP hydrolysis resulted in adenosine diphosphate (ADP) accumulation in the extracellular space. Pharmacological inhibition by ARL 67156 or gene silencing of the endogenous ecto-nucleoside triphosphate diphosphohydrolase (NTPDase) isoform 2 resulted in a 25% reduction in both ATP hydrolysis and ADP formation. Using intracellular calcium as a reporter, we found that although NTPDase2 hydrolyzed ATP and generated sustainable ADP levels, only ATP contributed to increased intracellular calcium via P2Y2 receptor activation. Furthermore, knocking down NTPDase2 potentiated the nanomolar ATP-induced intracellular calcium increase, suggesting that NTPDase2 globally attenuates nucleotide concentration in the pericellular microenvironment as well as locally shields receptors in the vicinity from being activated by extracellular ATP. Our findings reveal an important role of human keratinocyte NTPDase2 in modulating nucleotide signaling in the extracellular milieu of human epidermis.
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