Mull Makenzy L, Pratt Stephen J P, Thompson Keyata N, Annis David A, Lee Rachel M, Ju Julia A, Gilchrist Darin E, Stemberger Megan B, Boyman Liron, Lederer William J, Vitolo Michele I, Martin Stuart S
Graduate Program in Molecular Medicine, University of Maryland School of Medicine, 800 W. Baltimore St., Baltimore, MD 21201, USA.
Graduate Program in Biochemistry and Molecular Biology, University of Maryland School of Medicine, 108 N. Greene St., Baltimore, MD 21201, USA.
Int J Mol Sci. 2025 May 1;26(9):4286. doi: 10.3390/ijms26094286.
The tumor microenvironment and healing wounds both contain extremely high concentrations of adenosine triphosphate (ATP) compared to normal tissue. The P2Y2 receptor, an ATP-activated purinergic receptor, is typically associated with pulmonary, endothelial, and neurological cell signaling. Here, we examine ATP-dependent signaling in breast epithelial cells and how it is altered in metastatic breast cancer. Using rapid imaging techniques, we show how ATP-activated P2Y2 signaling causes an increase in intracellular Ca in non-tumorigenic breast epithelial cells, approximately 3-fold higher than their tumorigenic and metastatic counterparts. The non-tumorigenic cells respond to increased Ca with actin polymerization and localization to the cell edges after phalloidin staining, while the metastatic cells remain unaffected. The increase in intracellular Ca after ATP stimulation was blunted to control levels using a P2Y2 antagonist, which also prevented actin mobilization and significantly increased cell dissemination from spheroids in non-tumorigenic cells. Furthermore, the lack of Ca changes and actin mobilization in metastatic breast cancer cells could be due to the reduced P2Y2 expression, which correlates with poorer overall survival in breast cancer patients. This study elucidates the rapid changes that occur after elevated intracellular Ca in breast epithelial cells and how metastatic cancer cells have adapted to evade this cellular response.
与正常组织相比，肿瘤微环境和愈合伤口中的三磷酸腺苷（ATP）浓度极高。P2Y2受体是一种ATP激活的嘌呤能受体，通常与肺、内皮和神经细胞信号传导有关。在这里，我们研究乳腺上皮细胞中ATP依赖性信号传导以及它在转移性乳腺癌中是如何改变的。使用快速成像技术，我们展示了ATP激活的P2Y2信号传导如何导致非致瘤性乳腺上皮细胞内的Ca增加，比其致瘤性和转移性对应细胞高出约3倍。非致瘤性细胞在鬼笔环肽染色后，随着肌动蛋白聚合和定位到细胞边缘而对增加的Ca作出反应，而转移性细胞则不受影响。使用P2Y2拮抗剂将ATP刺激后细胞内Ca的增加减弱至对照水平，这也阻止了肌动蛋白的动员，并显著增加了非致瘤性细胞中球体的细胞扩散。此外，转移性乳腺癌细胞中缺乏Ca变化和肌动蛋白动员可能是由于P2Y2表达降低，这与乳腺癌患者较差的总体生存率相关。这项研究阐明了乳腺上皮细胞内Ca升高后发生的快速变化，以及转移性癌细胞如何适应以逃避这种细胞反应。
