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Effect of P2 receptor on the intracellular calcium increase by cancer cells in human umbilical vein endothelial cells

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Abstract

One of the important functions of vascular endothelial cells is as a barrier between blood and vascular tissue. This led us to speculate that cancer cells affect endothelial cells during metastasis. In the present study, we investigated the influence of human fibrosarcoma cells (HT-1080) on human umbilical vein endothelial cells (HUVEC), particularly intracellular calcium ion levels ([Ca2+]i), which are known to be an important intracellular signal transduction factor. HUVEC were treated with a fluorescent marker, and the fluorescence intensity of [Ca2+]i was then measured by phase contrast microscopic imaging. Extracellular adenosine triphosphate (ATP) release was measured using the chemiluminescence of luciferin–luciferase and a photon counting imaging system. HT-1080 (5 × 104 cells per dish) was found to increase [Ca2+]i in HUVEC. This [Ca2+]i rise was significantly reduced by U-73122 (phospholipase C inhibitor, 1 μM) and thapsigargin (calcium pump inhibitor, 1 μM). Interestingly, the [Ca2+]i rise in HUVEC was also significantly reduced by pyridoxalphosphare-6-azophenyl-2′, 4′-disulfonic acid, a P2Y receptor antagonist (100 μM) and apyrase, a nucleotidase inhibitor (2 U/ml). In addition, we observed ATP release from HT-1080. These results suggest that [Ca2+]i in HUVEC was increased through the phospholipase C-IP3 pathway via ATP release from cancer cells. We previously reported that extracellular ATP increased [Ca2+]i and enhanced macromolecular permeability via the P2Y receptor. In tumor metastasis, cancer cells may exploit these regulatory mechanisms in the endothelial cell layer.

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Acknowledgments

The authors would like to thank Ms. T. Tanaka, Ms. K. Nouda, Ms. S. Sato, and Ms. K. Motoya for their invaluable technical assistance. This research was supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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Authors and Affiliations

  1. Department of Pharmacology, School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women’s University, Nishinomiya, 663-8179, Japan

    Namie Nejime, Ryoko Yoshihara, Satomi Kagota, Noriko Yoshikawa, Kazuki Nakamura, Masaru Kunitomo & Kazumasa Shinozuka

  2. First Department of Pharmacology, School of Pharmaceutical Sciences, Kyushu University of Health and Welfare, Nobeoka, Miyazaki, 882-8508, Japan

    Naoko Tanaka

  3. Department of Environmental Physiology, Shimane University Faculty of Medicine, Izumo, 693-8501, Japan

    Michio Hashimoto

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  1. Namie Nejime
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Correspondence to Kazumasa Shinozuka.

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Nejime, N., Tanaka, N., Yoshihara, R. et al. Effect of P2 receptor on the intracellular calcium increase by cancer cells in human umbilical vein endothelial cells. Naunyn-Schmied Arch Pharmacol 377, 429–436 (2008). https://doi.org/10.1007/s00210-007-0259-2

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  • DOI: https://doi.org/10.1007/s00210-007-0259-2

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