ATP stimulates chemokine production via a store-operated calcium entry pathway in C6 glioma cells.
ATP stimulates chemokine production via a store-operated calcium entry pathway in C6 glioma cells.
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DOI：
10.1186/1471-2407-9-442
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发表时间：
2009-12-15
期刊：
影响因子：
3.8
通讯作者：
McLarnon JG
中科院分区：
文献类型：
--
作者：
Jantaratnotai N;Choi HB;McLarnon JG
胶质瘤是目前治疗最具挑战性的癌症之一，然而，对肿瘤细胞生物学的理解并不清楚。细胞外三磷酸腺苷（ATP）是调节肿瘤发生发展的重要信号分子。本研究探讨了药理学调节钙（Ca 2+）进入通过存储操作的通道（SOC）的细胞表达和生产的免疫细胞动员趋化因子在ATP刺激的C6胶质瘤细胞。用钙荧光分光光度法测定ATP刺激大鼠C6胶质瘤细胞后细胞内Ca ~（2+）[Ca ~（2+）]i的动员。用两种SOC抑制剂SKF 96365或钆预处理来检查对[Ca 2 +]i的影响。采用RT-PCR方法检测嘌呤能刺激对C6细胞代谢型P2 Y受体（P2 YR）、单核细胞趋化蛋白-1（MCP-1）和白细胞介素-8（IL-8）表达的影响。ELISA法检测ATP刺激胶质瘤细胞后MCP-1和IL-8的表达。将ATP（100 μM）应用于C6胶质瘤诱导[Ca 2 +]i增加，反应表现出两种衰减成分。在SOC抑制剂SKF 96365或钆存在下，或在无Ca 2+溶液中，ATP反应缺乏慢相，表明次要组分是由于SOC介导的Ca 2+内流。RT-PCR证实嘌呤能P2 Y亚型受体在C6细胞中的表达，这将作为SOC激活的前体。此外，ATP刺激的C6细胞显示趋化因子MCP-1和IL-8的表达增强，SKF 96365或钆有效地降低趋化因子的表达。还发现钆处理ATP刺激的C6细胞抑制MCP-1和IL-8的产生。这些结果表明ATP诱导的Ca 2+进入，介导的SOC在C6胶质瘤的激活，作为一种机制，导致增加细胞表达和释放的趋化因子。据预测，MCP-1和IL-8水平的升高会增强肿瘤细胞的移动性，并促进小胶质细胞向发育中的肿瘤中的募集，从而支持肿瘤生长。
Glioma present as one of the most challenging cancers to treat, however, understanding of tumor cell biology is not well understood. Extracellular adenosine triphosphate (ATP) could serve as a critical signaling molecule regulating tumor development. This study has examined pharmacological modulation of calcium (Ca2+) entry through store-operated channels (SOC) on cellular expression and production of immune-cell mobilizing chemokines in ATP-stimulated C6 glioma cells. Calcium spectrofluorometry was carried out to measure mobilization of intracellular Ca2+ [Ca2+]i following ATP stimulation of rat C6 glioma cells. Pretreatment with two inhibitors of SOC, SKF96365 or gadolinium, was used to examine for effects on [Ca2+]i. RT-PCR was performed to determine effects of purinergic stimulation on C6 cell expression of metabotropic P2Y receptors (P2YR) and the chemokines, monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8). ELISA was carried out to measure production of MCP-1 and IL-8 with ATP stimulation of glioma cells. Application of ATP (at 100 μM) to C6 glioma induced an increase in [Ca2+]i with the response exhibiting two components of decay. In the presence of the SOC inhibitors, SKF96365 or gadolinium, or with Ca2+-free solution, ATP responses lacked a slow phase suggesting the secondary component was due to SOC-mediated influx of Ca2+. RT-PCR confirmed expression of purinergic P2Y-subtype receptors in C6 cells which would serve as a precursor to activation of SOC. In addition, ATP-stimulated C6 cells showed enhanced expression of the chemokines, MCP-1 and IL-8, with SKF96365 or gadolinium effective in reducing chemokine expression. Gadolinium treatment of ATP-stimulated C6 cells was also found to inhibit the production of MCP-1 and IL-8. These results suggest ATP-induced Ca2+ entry, mediated by activation of SOC in C6 glioma, as a mechanism leading to increased cellular expression and release of chemokines. Elevated levels of MCP-1 and IL-8 are predicted to enhance the mobility of tumor cells and promote recruitment of microglia into developing tumors thereby supporting tumor growth.
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影响因子：
2.5
作者：
Badie, B;Schartner, J
通讯作者：
Schartner, J
影响因子：
11.2
作者：
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4.1
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5.3
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Andjelkovic, Anuska V.