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Disruption of P2Y2 Signaling Promotes Breast Tumor Cell Dissemination by Reducing ATP-Dependent Calcium Elevation and
Makenzy L Mull1, Stephen J P Pratt2, Keyata N Thompson3,4
1Graduate Program in Molecular Medicine, University of Maryland School of Medicine, 800 W. Baltimore St., Baltimore, MD 21201, USA.
Abstract:
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 Ca2+ in non-tumorigenic breast epithelial cells, approximately 3-fold higher than their tumorigenic and metastatic counterparts. The non-tumorigenic cells respond to increased Ca2+ with actin polymerization and localization to the cell edges after phalloidin staining, while the metastatic cells remain unaffected. The increase in intracellular Ca2+ 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 Ca2+ 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 Ca2+ in breast epithelial cells and how metastatic cancer cells have adapted to evade this cellular response.
Insights
Breast cancer cells show altered responses to adenosine triphosphate (ATP). Metastatic cells evade ATP-activated P2Y2 receptor signaling, impacting calcium (Ca2+) levels and cell behavior, unlike non-tumorigenic cells.
Area of Science:
- Cellular biology
- Cancer research
- Biochemistry
Background:
- High adenosine triphosphate (ATP) concentrations are found in tumor microenvironments and healing wounds.
- P2Y2 receptors, activated by ATP, mediate signaling in various cell types.
- ATP-dependent signaling in breast epithelial cells and its alteration in metastatic breast cancer are not fully understood.
Purpose of the Study:
- To investigate ATP-dependent P2Y2 receptor signaling in breast epithelial cells.
- To determine how this signaling pathway is altered in metastatic breast cancer.
- To elucidate the role of intracellular calcium (Ca2+) and actin dynamics in this process.
Main Methods:
- Utilized rapid imaging techniques to observe intracellular Ca2+ changes.
- Employed phalloidin staining to assess actin polymerization.
- Used a P2Y2 antagonist to block receptor activity.
- Analyzed cell dissemination from spheroids.
Main Results:
- Non-tumorigenic breast epithelial cells showed a 3-fold higher increase in intracellular Ca2+ upon ATP stimulation compared to metastatic cells.
- ATP-activated P2Y2 signaling induced actin polymerization at cell edges in non-tumorigenic cells, but not in metastatic cells.
- P2Y2 antagonism in non-tumorigenic cells reduced Ca2+ influx, prevented actin mobilization, and increased cell dissemination.
- Reduced P2Y2 expression in metastatic cells correlated with poorer patient survival.
Conclusions:
- Metastatic breast cancer cells exhibit a blunted response to ATP-activated P2Y2 signaling, characterized by impaired Ca2+ influx and actin mobilization.
- This evasion of cellular response may be linked to reduced P2Y2 receptor expression in metastatic cells.
- Findings highlight the role of P2Y2-mediated signaling in regulating breast epithelial cell behavior and suggest its potential as a therapeutic target.
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