title: Purine-metabolizing ectoenzymes control IL-8 production in human colon HT-29 cells.
abstract: 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.
authors.affiliation: Département de Microbiologie-Infectiologie et d'Immunologie, Faculté de Médecine, Université Laval, Québec, QC, Canada G1V 0A6 ; Centre de Recherche du CHU de Québec, 2705 Boulevard Laurier, Local T1-49, Québec, QC, Canada G1V 4G2.
authors.forename: Fariborz
authors.identifier: 0000-0001-9561-415X
authors.initials: F
authors.lastname: Bahrami
authors.affiliation: Département de Microbiologie-Infectiologie et d'Immunologie, Faculté de Médecine, Université Laval, Québec, QC, Canada G1V 0A6 ; Centre de Recherche du CHU de Québec, 2705 Boulevard Laurier, Local T1-49, Québec, QC, Canada G1V 4G2.
authors.forename: Filip
authors.initials: F
authors.lastname: Kukulski
authors.affiliation: Département de Microbiologie-Infectiologie et d'Immunologie, Faculté de Médecine, Université Laval, Québec, QC, Canada G1V 0A6 ; Centre de Recherche du CHU de Québec, 2705 Boulevard Laurier, Local T1-49, Québec, QC, Canada G1V 4G2.
authors.forename: Joanna
authors.identifier: 0000-0002-7127-4434
authors.initials: J
authors.lastname: Lecka
authors.affiliation: Centre de Recherche du CHU de Québec, 2705 Boulevard Laurier, Local T1-49, Québec, QC, Canada G1V 4G2.
authors.forename: Alain
authors.initials: A
authors.lastname: Tremblay
authors.affiliation: Centre de Recherche du CHU de Québec, 2705 Boulevard Laurier, Local T1-49, Québec, QC, Canada G1V 4G2.
authors.forename: Julie
authors.initials: J
authors.lastname: Pelletier
authors.affiliation: Département de Microbiologie-Infectiologie et d'Immunologie, Faculté de Médecine, Université Laval, Québec, QC, Canada G1V 0A6 ; Centre de Recherche du CHU de Québec, 2705 Boulevard Laurier, Local T1-49, Québec, QC, Canada G1V 4G2.
authors.forename: Liliana
authors.initials: L
authors.lastname: Rockenbach
authors.affiliation: Département de Microbiologie-Infectiologie et d'Immunologie, Faculté de Médecine, Université Laval, Québec, QC, Canada G1V 0A6 ; Centre de Recherche du CHU de Québec, 2705 Boulevard Laurier, Local T1-49, Québec, QC, Canada G1V 4G2.
authors.forename: Jean
authors.initials: J
authors.lastname: Sévigny
mesh_terms: D015720:5'-Nucleotidase / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000263:Adenylate Kinase / Q000378:metabolism; D019073:HT29 Cells; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011687:Purines / Q000378:metabolism*
chemical_list: D016209:Interleukin-8; D011687:Purines; D000263:Adenylate Kinase; D015720:5'-Nucleotidase; D000251:Adenosine Triphosphatases; C027201:ectoATPase; C030985:purine