HB-EGF Synthesis and Release Induced by Cholesterol Depletion of Human Epidermal Keratinocytes is Controlled by Extracellular ATP and Involves Both p38 and ERK1/2 Signaling Pathways
HB-EGF Synthesis and Release Induced by Cholesterol Depletion of Human Epidermal Keratinocytes is Controlled by Extracellular ATP and Involves Both p38 and ERK1/2 Signaling Pathways
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DOI：
10.1002/jcp.22496
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发表时间：
2011-06-01
影响因子：
5.6
通讯作者：
Poumay, Yves
中科院分区：
文献类型：
--
作者：
Giltaire, Severine;Lambert, Sylviane;Poumay, Yves
肝素结合的表皮生长因子样生长因子(HB-EGF)是一种自分泌/旁分泌的角质形成细胞生长因子，与表皮生长因子(EGF)受体家族结合，通过刺激角质形成细胞的增殖和迁移，在皮肤创面再上皮化过程中发挥重要作用。本研究采用甲基-β-环糊精(M-β-CD)去除胆固醇的方法，诱导人角质形成细胞自分泌的应激状态。MβCD可诱导HB-EGF的表达和释放。通过对细胞培养液的分析，发现M-β-CD作用1h后，细胞内可检测到大量的三磷酸腺苷。为了研究三磷酸腺苷是否参与HB-EGF的表达，用非水解性三磷酸腺苷类似物--三磷酸腺苷-伽马-S模拟细胞外释放的三磷酸腺苷。我们报道了用三磷酸腺苷-γ-S刺激角质形成细胞诱导HB-EGF的表达，并激活EGFR和ERK1/2。利用P2嘌呤能受体的拮抗剂，我们证明了脂筏破坏诱导的HB-EGF的合成依赖于ATP与P2嘌呤能受体的相互作用。此外，我们的数据表明，MAPKs p38和ERK1/2共同或独立地参与了HB-EGF基因表达的调节。这些发现为脂筏破坏后HB-EGF表达的信号通路提供了新的见解。综上所述，在脂筏破坏后，角质形成细胞释放大量的细胞外ATP。ATP通过与P2嘌呤能受体相互作用，并通过p38和ERK1/2信号途径诱导HB-EGF的合成和释放，以响应挑战环境。在角质形成细胞中，三磷酸腺苷的释放是一种早期的应激反应。J.细胞。物理。226：1651-1659,2011。(C)2010年Wiley-Liss公司
The heparin-binding EGF-like growth factor (HB-EGF) is an autocrine/paracrine keratinocyte growth factor, which binds to the epidermal growth factor (EGF) receptor family and plays a critical role during the re-epithelialization of cutaneous wound by stimulating the keratinocytes proliferation and migration. In this study, cellular stressing condition in autocrine cultures of human keratinocytes was induced by cholesterol depletion using methyl-beta-cyclodextrin (M beta CD). M beta CD treatment induces the expression and the release of HB-EGF. By analysis of the culture media, large amounts of cellular ATP were measured particularly after 1 h of M beta CD treatment. To investigate whether ATP contributes to the expression of HB-EGF, the nonhydrolyzable ATP analogue, ATP-gamma-S, was used to mimic the extracellular ATP released. We report that keratinocytes stimulated with ATP-gamma-S induce HB-EGF expression and activate EGFR and ERK1/2. Using an antagonist of P2 purinergic receptors, we demonstrate that HB-EGF synthesis induced by lipid rafts disruption is dependent on ATP interaction with P2 purinergic receptors. Moreover, our data suggest that both MAPKs p38 and ERK1/2 are involved together or independently in the regulation of HB-EGF gene expression. These findings provide new insight into the signaling pathway by which HB-EGF is expressed after lipid rafts disruption. In summary, after lipid raft disruption, keratinocytes release large amount of extracellular ATP. ATP induces HB-EGF synthesis and release by interacting with the P2 purinergic receptor and through p38 and ERK1/2 signaling in response to a challenging environment. A release of ATP acts as an early stress response in keratinocytes. J. Cell. Physiol. 226: 1651-1659, 2011. (C) 2010 Wiley-Liss, Inc.