Purine-Metabolizing Ectoenzymes Control IL-8 Production in Human Colon HT-29 Cells
Interleukin-8 (IL-8) plays key roles in both chronic inflammatory diseases and tumor modulation. We previously observed that IL-8 secretion and function can be modulated by nucleotide (P2) receptors. Here we investigated whether IL-8 release by intestinal epithelial HT-29 cells, a cancer cell line,...
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| Format: | Article |
| Language: | English |
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Wiley
2014-01-01
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| Series: | Mediators of Inflammation |
| Online Access: | http://dx.doi.org/10.1155/2014/879895 |
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| author | Fariborz Bahrami Filip Kukulski Joanna Lecka Alain Tremblay Julie Pelletier Liliana Rockenbach Jean Sévigny |
| author_facet | Fariborz Bahrami Filip Kukulski Joanna Lecka Alain Tremblay Julie Pelletier Liliana Rockenbach Jean Sévigny |
| author_sort | Fariborz Bahrami |
| collection | DOAJ |
| description | Interleukin-8 (IL-8) plays key roles in both chronic inflammatory diseases and tumor modulation. We previously observed that IL-8 secretion and function can be modulated by nucleotide (P2) receptors. Here we investigated whether IL-8 release by intestinal epithelial HT-29 cells, a cancer cell line, is modulated by extracellular nucleotide metabolism. We first identified that HT-29 cells regulated adenosine and adenine nucleotide concentration at their surface by the expression of the ectoenzymes NTPDase2, ecto-5′-nucleotidase, and adenylate kinase. The expression of the ectoenzymes was evaluated by RT-PCR, qPCR, and immunoblotting, and their activity was analyzed by RP-HPLC of the products and by detection of Pi produced from the hydrolysis of ATP, ADP, and AMP. In response to poly (I:C), with or without ATP and/or ADP, HT-29 cells released IL-8 and this secretion was modulated by the presence of NTPDase2 and adenylate kinase. Taken together, these results demonstrate the presence of 3 ectoenzymes at the surface of HT-29 cells that control nucleotide levels and adenosine production (NTPDase2, ecto-5′-nucleotidase and adenylate kinase) and that P2 receptor-mediated signaling controls IL-8 release in HT-29 cells which is modulated by the presence of NTPDase2 and adenylate kinase. |
| format | Article |
| id | doaj-art-fdd9b8806b3a411986ba8557bf6fb0d2 |
| institution | Kabale University |
| issn | 0962-9351 1466-1861 |
| language | English |
| publishDate | 2014-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Mediators of Inflammation |
| spelling | doaj-art-fdd9b8806b3a411986ba8557bf6fb0d22025-02-03T00:59:54ZengWileyMediators of Inflammation0962-93511466-18612014-01-01201410.1155/2014/879895879895Purine-Metabolizing Ectoenzymes Control IL-8 Production in Human Colon HT-29 CellsFariborz Bahrami0Filip Kukulski1Joanna Lecka2Alain Tremblay3Julie Pelletier4Liliana Rockenbach5Jean Sévigny6Département de Microbiologie-Infectiologie et d’Immunologie, Faculté de Médecine, Université Laval, Québec, QC, G1V 0A6, CanadaDépartement de Microbiologie-Infectiologie et d’Immunologie, Faculté de Médecine, Université Laval, Québec, QC, G1V 0A6, CanadaDépartement de Microbiologie-Infectiologie et d’Immunologie, Faculté de Médecine, Université Laval, Québec, QC, G1V 0A6, CanadaCentre de Recherche du CHU de Québec, 2705 Boulevard Laurier, Local T1-49, Québec, QC, G1V 4G2, CanadaCentre de Recherche du CHU de Québec, 2705 Boulevard Laurier, Local T1-49, Québec, QC, G1V 4G2, CanadaDépartement de Microbiologie-Infectiologie et d’Immunologie, Faculté de Médecine, Université Laval, Québec, QC, G1V 0A6, CanadaDépartement de Microbiologie-Infectiologie et d’Immunologie, Faculté de Médecine, Université Laval, Québec, QC, G1V 0A6, CanadaInterleukin-8 (IL-8) plays key roles in both chronic inflammatory diseases and tumor modulation. We previously observed that IL-8 secretion and function can be modulated by nucleotide (P2) receptors. Here we investigated whether IL-8 release by intestinal epithelial HT-29 cells, a cancer cell line, is modulated by extracellular nucleotide metabolism. We first identified that HT-29 cells regulated adenosine and adenine nucleotide concentration at their surface by the expression of the ectoenzymes NTPDase2, ecto-5′-nucleotidase, and adenylate kinase. The expression of the ectoenzymes was evaluated by RT-PCR, qPCR, and immunoblotting, and their activity was analyzed by RP-HPLC of the products and by detection of Pi produced from the hydrolysis of ATP, ADP, and AMP. In response to poly (I:C), with or without ATP and/or ADP, HT-29 cells released IL-8 and this secretion was modulated by the presence of NTPDase2 and adenylate kinase. Taken together, these results demonstrate the presence of 3 ectoenzymes at the surface of HT-29 cells that control nucleotide levels and adenosine production (NTPDase2, ecto-5′-nucleotidase and adenylate kinase) and that P2 receptor-mediated signaling controls IL-8 release in HT-29 cells which is modulated by the presence of NTPDase2 and adenylate kinase.http://dx.doi.org/10.1155/2014/879895 |
| spellingShingle | Fariborz Bahrami Filip Kukulski Joanna Lecka Alain Tremblay Julie Pelletier Liliana Rockenbach Jean Sévigny Purine-Metabolizing Ectoenzymes Control IL-8 Production in Human Colon HT-29 Cells Mediators of Inflammation |
| title | Purine-Metabolizing Ectoenzymes Control IL-8 Production in Human Colon HT-29 Cells |
| title_full | Purine-Metabolizing Ectoenzymes Control IL-8 Production in Human Colon HT-29 Cells |
| title_fullStr | Purine-Metabolizing Ectoenzymes Control IL-8 Production in Human Colon HT-29 Cells |
| title_full_unstemmed | Purine-Metabolizing Ectoenzymes Control IL-8 Production in Human Colon HT-29 Cells |
| title_short | Purine-Metabolizing Ectoenzymes Control IL-8 Production in Human Colon HT-29 Cells |
| title_sort | purine metabolizing ectoenzymes control il 8 production in human colon ht 29 cells |
| url | http://dx.doi.org/10.1155/2014/879895 |
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