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Growth inhibition of breast cancer cells induced by exogenous ATP
1Department of Cell Research and Immunology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Israel.
Abstract:
Addition of ATP (> 0.1 mM) to cultures of human breast cancer T47D cells resulted in an inhibition of cell proliferation. The inhibition was found to be specific for ATP, and dependent on its concentration. Growth inhibition continued for at least three days, although ATP and its hydrolysis products were metabolized within one day. Conditioned medium from ATP-treated cultures (CM+) was found to inhibit the growth of cells that were not exposed to ATP. This is an indication that extracellular factors, besides ATP, are involved in the inhibition process. The inhibition was maintained after dialysis of the CM+, using an 8 kDa cut-off membrane. Conditioned medium from untreated cultures (CM-), however, only slightly affected cell growth. The data suggest that the CM(+)-induced cell growth inhibition is mediated by an ATP-activated growth inhibiting factor. Flow microfluorometry and thymidine incorporation experiments have shown that the growth arrest is mainly due to the elongation of the S-phase of the cell cycle.
Insights
Adenosine triphosphate (ATP) inhibits human breast cancer cell growth. An ATP-activated extracellular factor in conditioned medium mediates this inhibition, causing cell cycle arrest.
Area of Science:
- Cancer Research
- Cell Biology
- Molecular Pharmacology
Background:
- Extracellular nucleotides, including adenosine triphosphate (ATP), play diverse roles in cell signaling.
- The specific impact of exogenous ATP on human breast cancer cell proliferation requires further elucidation.
Purpose of the Study:
- To investigate the effect of ATP on the proliferation of human breast cancer T47D cells.
- To identify potential extracellular factors mediating ATP-induced growth inhibition.
Main Methods:
- Treatment of T47D cell cultures with varying concentrations of ATP.
- Analysis of cell proliferation rates.
- Preparation and testing of conditioned medium (CM+) from ATP-treated cells.
- Dialysis of CM+ using an 8 kDa cut-off membrane.
- Flow microfluorometry and thymidine incorporation assays to assess cell cycle progression.
Main Results:
- ATP significantly inhibited T47D cell proliferation in a dose-dependent manner.
- Conditioned medium from ATP-treated cultures (CM+) inhibited growth of untreated cells, indicating an extracellular mediator.
- The inhibitory effect of CM+ persisted after dialysis, suggesting a stable factor.
- Cell cycle analysis revealed that ATP-induced growth arrest primarily results from S-phase elongation.
Conclusions:
- ATP acts as a specific inhibitor of human breast cancer T47D cell proliferation.
- An ATP-activated extracellular factor, stable and with a molecular weight > 8 kDa, mediates the observed growth inhibition.
- The mechanism involves a delay in cell cycle progression, specifically an extended S-phase.