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Carcinoma-specific expression of P2Y(11) receptor and its contribution in ATP-induced purinergic signalling and cell migration in human hepatocellular carcinoma cells

Extracellular ATP-induced Ca(2+) signalling is critical in regulating diverse physiological and disease processes. Emerging evidence suggests high concentrations of extracellular ATP in tumour tissues. In this study, we examined the P2 receptor for ATP-induced Ca(2+) signalling in human hepatocellul...

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Autores principales: Khalid, Madiha, Brisson, Lucie, Tariq, Menahil, Hao, Yunjie, Guibon, Roseline, Fromont, Gaëlle, Syed Mortadza, Sharifah Alawieyah, Mousawi, Fatema, Manzoor, Sobia, Roger, Sébastien, Jiang, Lin-Hua
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Impact Journals LLC 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5514908/
https://www.ncbi.nlm.nih.gov/pubmed/28418839
http://dx.doi.org/10.18632/oncotarget.16191
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author Khalid, Madiha
Brisson, Lucie
Tariq, Menahil
Hao, Yunjie
Guibon, Roseline
Fromont, Gaëlle
Syed Mortadza, Sharifah Alawieyah
Mousawi, Fatema
Manzoor, Sobia
Roger, Sébastien
Jiang, Lin-Hua
author_facet Khalid, Madiha
Brisson, Lucie
Tariq, Menahil
Hao, Yunjie
Guibon, Roseline
Fromont, Gaëlle
Syed Mortadza, Sharifah Alawieyah
Mousawi, Fatema
Manzoor, Sobia
Roger, Sébastien
Jiang, Lin-Hua
author_sort Khalid, Madiha
collection PubMed
description Extracellular ATP-induced Ca(2+) signalling is critical in regulating diverse physiological and disease processes. Emerging evidence suggests high concentrations of extracellular ATP in tumour tissues. In this study, we examined the P2 receptor for ATP-induced Ca(2+) signalling in human hepatocellular carcinoma (HCC) cells. Fura-2-based measurements of the intracellular Ca(2+) concentration ([Ca(2+)](i)) showed that extracellular ATP induced an increase in the [Ca(2+)](i) in human HCC Huh-7 and HepG2 cells. NF546, a P2Y(11) receptor agonist was equally effective in inducing an increase in the [Ca(2+)](i). In contrast, agonists for the P2X receptors (αβmeATP and BzATP), P2Y(1) receptor (MRS2365) or P2Y(2) receptor (MRS2768) were ineffective. In addition, ATP/NF546-induced increases in the [Ca(2+)](i) were strongly inhibited by treatment with NF340, a P2Y(11) receptor antagonist. Immunofluorescent confocal imaging and western blotting analysis consistently demonstrated the P2Y(11) receptor expression in Huh-7 and HepG2 cells. Transfection with P2Y(11)-specific siRNA attenuated the P2Y(11) receptor protein expression level and also reduced NF546-induced increase in the [Ca(2+)](i). Importantly, immunohistochemistry revealed that the P2Y(11) receptor was expressed at very high level in human HCC tissues and, by contrast, it was barely detected in normal liver tissues. Trans-well cell migration assay demonstrated that ATP and NF546 induced concentration-dependent stimulation of Huh-7 cell migration. Treatment with NF340 prevented ATP-induced stimulation of cell migration. Taken together, our results show carcinoma-specific expression of the P2Y(11) receptor and its critical role in mediating ATP-inducing Ca(2+) signalling and regulating cell migration in human HCC cells.
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spelling pubmed-55149082017-07-24 Carcinoma-specific expression of P2Y(11) receptor and its contribution in ATP-induced purinergic signalling and cell migration in human hepatocellular carcinoma cells Khalid, Madiha Brisson, Lucie Tariq, Menahil Hao, Yunjie Guibon, Roseline Fromont, Gaëlle Syed Mortadza, Sharifah Alawieyah Mousawi, Fatema Manzoor, Sobia Roger, Sébastien Jiang, Lin-Hua Oncotarget Research Paper Extracellular ATP-induced Ca(2+) signalling is critical in regulating diverse physiological and disease processes. Emerging evidence suggests high concentrations of extracellular ATP in tumour tissues. In this study, we examined the P2 receptor for ATP-induced Ca(2+) signalling in human hepatocellular carcinoma (HCC) cells. Fura-2-based measurements of the intracellular Ca(2+) concentration ([Ca(2+)](i)) showed that extracellular ATP induced an increase in the [Ca(2+)](i) in human HCC Huh-7 and HepG2 cells. NF546, a P2Y(11) receptor agonist was equally effective in inducing an increase in the [Ca(2+)](i). In contrast, agonists for the P2X receptors (αβmeATP and BzATP), P2Y(1) receptor (MRS2365) or P2Y(2) receptor (MRS2768) were ineffective. In addition, ATP/NF546-induced increases in the [Ca(2+)](i) were strongly inhibited by treatment with NF340, a P2Y(11) receptor antagonist. Immunofluorescent confocal imaging and western blotting analysis consistently demonstrated the P2Y(11) receptor expression in Huh-7 and HepG2 cells. Transfection with P2Y(11)-specific siRNA attenuated the P2Y(11) receptor protein expression level and also reduced NF546-induced increase in the [Ca(2+)](i). Importantly, immunohistochemistry revealed that the P2Y(11) receptor was expressed at very high level in human HCC tissues and, by contrast, it was barely detected in normal liver tissues. Trans-well cell migration assay demonstrated that ATP and NF546 induced concentration-dependent stimulation of Huh-7 cell migration. Treatment with NF340 prevented ATP-induced stimulation of cell migration. Taken together, our results show carcinoma-specific expression of the P2Y(11) receptor and its critical role in mediating ATP-inducing Ca(2+) signalling and regulating cell migration in human HCC cells. Impact Journals LLC 2017-03-14 /pmc/articles/PMC5514908/ /pubmed/28418839 http://dx.doi.org/10.18632/oncotarget.16191 Text en Copyright: © 2017 Khalid et al. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/) (CC-BY), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Paper
Khalid, Madiha
Brisson, Lucie
Tariq, Menahil
Hao, Yunjie
Guibon, Roseline
Fromont, Gaëlle
Syed Mortadza, Sharifah Alawieyah
Mousawi, Fatema
Manzoor, Sobia
Roger, Sébastien
Jiang, Lin-Hua
Carcinoma-specific expression of P2Y(11) receptor and its contribution in ATP-induced purinergic signalling and cell migration in human hepatocellular carcinoma cells
title Carcinoma-specific expression of P2Y(11) receptor and its contribution in ATP-induced purinergic signalling and cell migration in human hepatocellular carcinoma cells
title_full Carcinoma-specific expression of P2Y(11) receptor and its contribution in ATP-induced purinergic signalling and cell migration in human hepatocellular carcinoma cells
title_fullStr Carcinoma-specific expression of P2Y(11) receptor and its contribution in ATP-induced purinergic signalling and cell migration in human hepatocellular carcinoma cells
title_full_unstemmed Carcinoma-specific expression of P2Y(11) receptor and its contribution in ATP-induced purinergic signalling and cell migration in human hepatocellular carcinoma cells
title_short Carcinoma-specific expression of P2Y(11) receptor and its contribution in ATP-induced purinergic signalling and cell migration in human hepatocellular carcinoma cells
title_sort carcinoma-specific expression of p2y(11) receptor and its contribution in atp-induced purinergic signalling and cell migration in human hepatocellular carcinoma cells
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5514908/
https://www.ncbi.nlm.nih.gov/pubmed/28418839
http://dx.doi.org/10.18632/oncotarget.16191
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