Emodin Inhibits ATP-Induced Proliferation and Migration by Suppressing P2Y Receptors in Human Lung Adenocarcinoma Cells
Emodin Inhibits ATP-Induced Proliferation and Migration by Suppressing P2Y Receptors in Human Lung Adenocarcinoma Cells
复制标题
大黄素通过抑制人肺腺癌细胞中的 P2Y 受体来抑制 ATP 诱导的增殖和迁移
DOI：
10.1159/000485495
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发表时间：
2017-01-01
影响因子：
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通讯作者：
Shen, Linlin
中科院分区：
文献类型：
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作者：
Wang, Xia;Li, Long;Shen, Linlin
背景/目的：细胞外ATP通过激活细胞表面的P2受体发挥多种重要功能。P2Y受体在人肺腺癌细胞(A549细胞)的三磷酸腺苷诱导反应中起重要作用。大黄素是从大黄中分离得到的一种具有抗癌活性的大黄素类化合物。在本研究中，我们通过抑制A549细胞中依赖于P2Y受体的钙离子浓度和核因子-κB信号通路，观察了大黄素对A549细胞的增殖、迁移和上皮-间充质转化的抑制作用。方法：用大黄素预先处理A549细胞，然后用三磷酸腺苷刺激A549细胞。用Fluo-8/AM染色检测细胞内钙离子浓度([Ca~(2+)]i)。用CCK8法和流式细胞仪检测细胞增殖和细胞周期进程，伤口愈合和免疫印迹检测细胞迁移和相关蛋白(bcl2、bax、claudin-1、NF-κB)的表达。结果：大黄素呈浓度依赖性拮抗三磷酸腺苷/尿苷三磷酸诱导的细胞[Ca~(2+)]i升高和细胞增殖，同时减少三磷酸腺苷诱导的细胞在S时的积聚。此外，大黄素改变了Bcl2、Bax和claudin-1的蛋白丰度，并减弱了ATP引起的EMT。这种由ATP诱导的细胞反应也被非选择性的P2Y受体拮抗剂苏拉明抑制，其方式与大黄素类似。此外，大黄素还可抑制NF-κB的激活，从而抑制其诱导的增殖、迁移和内皮细胞转化。结论：大黄素通过抑制P2Y受体介导的[Ca~(2+)]i升高和核转录因子-κB信号通路，抑制三磷酸腺苷诱导的A549细胞的增殖、迁移和内胚层转化。
Background/Aims: Extracellular ATP performs multiple important functions via activation of P2 receptors on the cell surface. P2Y receptors play critical roles in ATP evoked response in human lung adenocarcinoma cells (A549 cells). Emodin is an anthraquinone derivative originally isolated from Chinese rhubarb, possesses anticancer properties. In this study we examined the inhibiting effects of emodin on proliferation, migration and epithelial-mesenchymal transition (EMT) by suppressing P2Y receptors-dependent Ca2+ increase and nuclear factor-κB (NF-KB) signaling in A549 cells. Methods: A549 cells were pretreated with emodin before stimulation with ATP for the indicated time. Then, intracellular Ca2+ concentration ([Ca2+]i) was measured by Fluo-8/AM staining. Cell proliferation and cell cycle progression were tested by CCK8 assay and flow cytometry In addition, wound healing and western blot were performed to determine cell migration and related protein levels (Bcl-2, Bax, claudin-1, NF-κB). Results: Emodin blunted ATP/UTP-induced increase of [Ca2+]i and cell proliferation concentration-dependently Meanwhile, it decreased ATP-induced cells accumulation in the S phase. Furthermore, emodin altered protein abundance of Bcl-2, Bax and claudin-1 and attenuated EMT caused by ATP. Such ATP-induced cellular reactions were also inhibited by a nonselective P2Y receptors antagonist, suramin, in a similar way to emodin. Besides, emodin could inhibit activation of NF-κB, thus suppressed ATP-induced proliferation, migration and EMT. Conclusion: Our results demonstrated that emodin inhibits ATP-induced proliferation, migration, EMT by suppressing P2Y receptors-mediated [Ca2+]i increase and NF-κB signaling in A549 cells.